Peptides and peptide mimetics to treat pathologies characterized by an inflammatory response

ABSTRACT

This invention provides novel active agents (e.g. peptides, small organic molecules, amino acid pairs, etc.) peptides that ameliorate one or more symptoms of atherosclerosis and/or other pathologies characterized by an inflammatory response. In certain embodiment, the peptides resemble a G* amphipathic helix of apolipoprotein J. The agents are highly stable and readily administered via an oral route.

This application is a continuation of and claims priority from U.S. Ser.No. 11/407,390, filed on Apr. 18, 2006, which claims priority to andbenefit of U.S. Ser. No. 60/697,495, filed Jul. 7, 2005 and to U.S. Ser.No. 60/676,431 filed on Apr. 29, 2005, both of which are incorporatedherein by reference in their entirety for all purposes. This applicationis also a Continuation-in-Part of U.S. Ser. No. 10/423,830, filed onApr. 25, 2003, now issued U.S. Pat. No. 7,199,102, which is aContinuation-in-Part of U.S. Ser. No. 10/273,386, filed on Oct. 16,2002, now issued U.S. Pat. No. 7,166,578, which is aContinuation-in-Part of U.S. Ser. No. 10/187,215, filed on Jun. 28,2002, now issued U.S. Pat. No. 7,144,862, which is aContinuation-in-Part of U.S. Ser. No. 09/896,841, filed on Jun. 29,2001, now issued U.S. Pat. No. 6,933,279, which is aContinuation-in-Part of U.S. Ser. No. 09/645,454, filed on Aug. 24,2000, now issued U.S. Pat. No. 6,664,230, all of which are incorporatedherein by reference in their entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

This work was supported, in part, by Grant No: HL30568 from the NationalHeart Blood Lung Institute of the National Institutes of Health. TheGovernment of the United States of America may have certain rights inthis invention.

FIELD OF THE INVENTION

This invention relates to the field of atherosclerosis and otherconditions characterized by inflammation and/or the formation of variousoxidized species. In particular, this invention pertains to theidentification of classes of active agents that are orally administrableand that ameliorate one or more symptoms of conditions characterized byan inflammatory response and/or the formation of various oxidizedspecies.

BACKGROUND OF THE INVENTION

The introduction of statins (e.g., Mevacor®, Lipitor®, etc.) has reducedmortality from heart attack and stroke by about one-third. However,heart attack and stroke remain the major cause of death and disability,particularly in the United States and in Western European countries.Heart attack and stroke are the result of a chronic inflammatorycondition, which is called atherosclerosis.

Several causative factors are implicated in the development ofcardiovascular disease including hereditary predisposition to thedisease, gender, lifestyle factors such as smoking and diet, age,hypertension, and hyperlipidemia, including hypercholesterolemia.Several of these factors, particularly hyperlipidemia andhypercholesteremia (high blood cholesterol concentrations) provide asignificant risk factor associated with atherosclerosis.

Cholesterol is present in the blood as free and esterified cholesterolwithin lipoprotein particles, commonly known as chylomicrons, very lowdensity lipoproteins (VLDLs), low density lipoproteins (LDLs), and highdensity lipoproteins (HDLs). Concentration of total cholesterol in theblood is influenced by (1) absorption of cholesterol from the digestivetract, (2) synthesis of cholesterol from dietary constituents such ascarbohydrates, proteins, fats and ethanol, and (3) removal ofcholesterol from blood by tissues, especially the liver, and subsequentconversion of the cholesterol to bile acids, steroid hormones, andbiliary cholesterol.

Maintenance of blood cholesterol concentrations is influenced by bothgenetic and environmental factors. Genetic factors include concentrationof rate-limiting enzymes in cholesterol biosynthesis, concentration ofreceptors for low density lipoproteins in the liver, concentration ofratelimiting enzymes for conversion of cholesterols bile acids, rates ofsynthesis and secretion of lipoproteins and gender of person.Environmental factors influencing the hemostasis of blood cholesterolconcentration in humans include dietary composition, incidence ofsmoking, physical activity, and use of a variety of pharmaceuticalagents. Dietary variables include the amount and type of fat (saturatedand polyunsaturated fatty acids), the amount of cholesterol, amount andtype of fiber, and perhaps the amounts of vitamins such as vitamin C andD and minerals such as calcium.

Low density lipoprotein (LDL) oxidation has been strongly implicated inthe pathogenesis of atherosclerosis. High density lipoprotein (HDL) hasbeen found to be capable of protecting against LDL oxidation, but insome instances has been found to accelerate LDL oxidation. Importantinitiating factors in atherosclerosis include the production ofLDL-derived oxidized phospholipids.

Normal HDL has the capacity to prevent the formation of these oxidizedphospholipids and also to inactivate these oxidized phospholipids oncethey have formed. However, under some circumstances HDL can be convertedfrom an anti-inflammatory molecule to a pro-inflammatory molecule thatactually promotes the formation of these oxidized phospholipids.

It has been suggested that HDL and LDL function as part of the innateimmune system (Navab et al. (2001) Arterioscler. Thromb. Vasc. Biol.,21: 481-488). The generation of anti-inflammatory HDL has been achievedusing class A amphipathic helical peptides that mimic the major proteinof HDL, apolipoprotein A-I (apo A-I) (see, e.g., WO 02/15923).

SUMMARY OF THE INVENTION

This invention provides novel compositions and methods to ameliorate oneor more symptoms of a vascular condition and/or a conditioncharacterized by an inflammatory response and/or a conditioncharacterized by the formation of oxidized reactive species in a mammal.

Thus, in certain embodiments, this invention provides a peptide thatameliorates a symptom of atherosclerosis, where the peptide comprisesthe amino acid sequence or the retro amino acid sequence of a peptidelisted in Table 6. In another embodiment this invention provides apeptide that ameliorates a symptom of atherosclerosis, where thepeptide: consists of 18 amino acids, the 18 amino acids consisting of 3alanines (A), 2 aspartates (D), 2 glutamates (E), 4 phenylalanines (F),4 lysines (K), 1 valine (V), 1 tryptophan (W), and 1 tyrosine (Y); wherethe peptide forms a class A amphipathic helix; comprises at least one“D” amino acid residue; and protects a phospholipid against oxidation byan oxidizing agent. In certain embodiments these peptides include butare not limited to a peptide having the amino acid sequence or the retroamino acid sequence of a peptide listed in Table 4. In still anotherembodiment, this invention provides a peptide that ameliorates a symptomof atherosclerosis, where the peptide: ranges in length from about 18 to37 amino acids and comprises at least 3 alanines (A), 2 aspartates (D),2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine (V), 1tryptophan (W), 1 tyrosine (Y); where the peptide forms a class Aamphipathic helix; comprises at least one “D” amino acid residue; andprotects a phospholipid against oxidation by an oxidizing agent. Incertain embodiments these peptides comprise an amino acid sequenceselected from the group consisting ofD-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F (SEQ ID NO: 1179),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1180),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F(SEQ ID NO: 1181),-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1182),D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-P-D-K-L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-F(SEQ ID NO: 1183),D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-P-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L(SEQ ID NO: 1184),D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-(SEQ ID NO: 1185), or the reverse of any of these sequences. In stillyet another embodiment this invention provides a peptide that forms aclass A amphipathic helix or a class Y amphipathic helix and isdescribed by the formula:D¹-X¹-X²-K¹-Y¹-X³-X⁴-D²-K²-X⁵-Y-D³-K³-X⁶-K⁴-D⁴-Y²-X⁷, where X¹, X², X³,X⁴, X⁵, and X⁶ are independently selected from the group consisting ofLeu, norLeu, Val, Ile, Trp, Phe, Tyr, β-Nal, and α-Nal, and all Xresidues are on the non-polar face of the peptide, except for one thatcan be on the polar face between two K residues; K¹, K², K³, and K⁴ areindependently Lys or Arg, and no more than two K's are adjacent to eachother in a helical wheel diagram of the peptide; Y¹ and Y² areindependently selected from the group consisting of Ala, His, Ser, Gln,Asn, and Thr, when present on the non-polar face of the molecule; whenone of Y¹ or Y² are present on the polar face of the molecule, the Y¹ orY² on the polar face of the molecule is selected from the groupconsisting of Ala, His, Ser, Gln, Asn, and Thr; D¹, D², D³, and D⁴ areindependently Asp or Glu, and no more than 3 Ds are contiguous in ahelical wheel diagram of the peptide, and the remaining D is separatedfrom the other D's by a Y. In certain embodiments these peptidescomprise the amino acid sequence or the retro amino acid sequence of apeptide listed in Table 5.

In certain embodiments any one or more of these peptides furthercomprise a protecting group coupled to the amino or carboxyl terminus.In certain embodiments the peptides comprise a first protecting groupcoupled to the amino terminus and a second protecting group coupled tothe carboxyl terminus. In certain embodiments the protecting groups canbe independently selected from the group consisting of acetyl, amide,and 3 to 20 carbon alkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group,1-fluorenecarboxylic group, 9-florenecarboxylic group,9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan),Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt),4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl(Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethylchroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl(MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz),3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z),2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).

In certain embodiments the peptide comprises a protecting group coupledto the amino terminal and the amino terminal protecting group is aprotecting group selected from the group consisting of acetyl,propeonyl, and a 3 to 20 carbon alkyl. In certain embodiments thepeptide comprises a protecting group coupled to the carboxyl terminaland the carboxyl terminal protecting group is an amide. In certainembodiments the peptide comprises: a first protecting group coupled tothe amino terminus where the protecting group is a protecting groupselected from the group consisting of acetyl, propeonyl, and a 3 to 20carbon alkyl; and a second protecting group coupled to the carboxylterminal and the carboxyl terminal protecting group is an amide.

In various embodiments one or more amino acids comprising the peptideare “D” amino acids. In various embodiments all amino acids comprisingthe peptide “D” amino acids. The peptide(s) can, optionally, bemixed/combined with a pharmacologically acceptable excipient. In certainembodiments the excipient is an excipient suitable for oraladministration to a mammal.

In certain embodiments this invention provides methods of treating avascular condition and/or a condition characterized by an inflammatoryresponse and/or a condition characterized by the formation of oxidizedreactive species in a mammal. The methods typically involveadministering to a mammal in need thereof one or more of the activeagents described in Tables 2-18, and/or a small organic molecule asdescribed herein in an amount sufficient to ameliorate one or moresymptoms of the condition. In certain embodiments the active agent is apolypeptide comprising the amino acid sequence of 4F (SEQ ID NO:5). Incertain embodiments the administration is by a route selected from thegroup consisting of oral administration, nasal administration, rectaladministration, intraperitoneal injection, and intravascular injection,subcutaneous injection, transcutaneous administration, and intramuscularinjection. In certain embodiments the active agent is administered inconjunction with a drug selected from the group consisting of CETPinhibitors, FTY720, Certican, DPP4 inhibitors, Calcium channel blockers,ApoA1 derivative or mimetic or agonist, PPAR agonists, Steroids,Gleevec, Cholesterol Absorption blockers (Zetia), Vytorin, Any ReninAngiotensin pathway blockers, Angiotensin II receptor antagonist (Diovanetc), ACE inhibitors, Renin inhibitors, MR antagonist and Aldosteronesynthase inhibitor, Beta-blockers, Alpha-adrenergic antagonists, LXRagonist, FXR agonist, Scavenger Receptor B1 agonist, ABCA1 agonist,Adiponectic receptor agonist or adiponectin inducers, Stearoyl-CoADesaturase I (SCD1) inhibitor, Cholesterol synthesis inhibitors(non-statins), Diacylglycerol Acyltransferase I (DGAT1) inhibitor,Acetyl CoA Carboxylase 2 inhibitor, PAI-1 inhibitor, LP-PLA2 inhibitor,GLP-1, Glucokinase activator, CB-1 agonist, AGE inhibitoribreaker, PKCinhibitors, Anti-thrombotic/coagulants: Aspirin, ADP receptor blockerse.g. Clopidigrel, Factor Xa inhibitor, GPIIb/IIIa inhibitor, Factor VIIainhibitor, Warfarin, Low molecular weight heparin, Tissue factorinhibitor, Anti-inflammatory drugs: Probucol and derivative e.g.AGI-1067 etc, CCR2 antagonist, CX3CR1 antagonist, IL-1 antagonist,Nitrates and NO donors, and Phosphodiesterase inhibitors.

In various embodiments this invention provides for the use of an activeagent described in Tables 2-18, and/or a small organic molecule asdescribed herein in a treatment of a condition selected from the groupconsisting of atherosclerotic plaque formation, atherosclerotic lesionformation, myocardial infarction, stroke, congestive heart failure,arteriole function, arteriolar disease, arteriolar disease associatedwith aging, arteriolar disease associated with Alzheimer's disease,arteriolar disease associated with chronic kidney disease, arteriolardisease associated with hypertension, arteriolar disease associated withmulti-infarct dementia, arteriolar disease associated with subarachnoidhemorrhage, peripheral vascular disease, chronic obstructive pulmonarydisease (COPD), emphysema, asthma, idiopathic pulmonary fibrosis,pulmonary fibrosis, adult respiratory distress syndrome, osteoporosis,Paget's disease, coronary calcification, rheumatoid arthritis,polyarteritis nodosa, polymyalgia rheumatica, lupus erythematosus,multiple sclerosis, Wegener's granulomatosis, central nervous systemvasculitis (CNSV), Sjögren's syndrome, scleroderma, polymyositis, AIDSinflammatory response, bacterial infection, fungal infection, viralinfection, parasitic infection, influenza, avian flu, viral pneumonia,endotoxic shock syndrome, sepsis, sepsis syndrome, trauma/wound, organtransplant, transplant atherosclerosis, transplant rejection, cornealulcer, chronic/non-healing wound, ulcerative colitis, reperfusion injury(prevent and/or treat), ischemic reperfusion injury (prevent and/ortreat), spinal cord injuries (mitigating effects), cancers,myeloma/multiple myeloma, ovarian cancer, breast cancer, colon cancer,bone cancer osteoarthritis, inflammatory bowel disease, allergicrhinitis, cachexia, diabetes, Alzheimer's disease, implanted prosthesis,biofilm formation, Crohns' disease, dermatitis, acute and chronic,eczema, psoriasis, contact dermatitis, scleroderma, Type I Diabetes,Type II Diabetes, juvenile onset diabetes, prevention of the onset ofdiabetes, diabetic nephropathy, diabetic neuropathy, diabeticretinopathy, erectile dysfunction, macular degeneration, multiplesclerosis, nephropathy, neuropathy, Parkinson's Disease, peripheralvascular disease, and meningitis.

This invention additionally provides for the use of active agentdescribed in Tables 2-18, and/or a small organic molecule as describedherein for the manufacture of a medicament for the treatment of acondition selected from the group consisting of atherosclerotic plaqueformation, atherosclerotic lesion formation, myocardial infarction,stroke, congestive heart failure, arteriole function, arteriolardisease, arteriolar disease associated with aging, arteriolar diseaseassociated with Alzheimer's disease, arteriolar disease associated withchronic kidney disease, arteriolar disease associated with hypertension,arteriolar disease associated with multi-infarct dementia, arteriolardisease associated with subarachnoid hemorrhage, peripheral vasculardisease, chronic obstructive pulmonary disease (COPD), emphysema,asthma, idiopathic pulmonary fibrosis, pulmonary fibrosis, adultrespiratory distress syndrome, osteoporosis, Paget's disease, coronarycalcification, rheumatoid arthritis, polyarteritis nodosa, polymyalgiarheumatica, lupus erythematosus, multiple sclerosis, Wegener'sgranulomatosis, central nervous system vasculitis (CNSV), Sjögren'ssyndrome, scleroderma, polymyositis, AIDS inflammatory response,bacterial infection, fungal infection, viral infection, parasiticinfection, influenza, avian flu, viral pneumonia, endotoxic shocksyndrome, sepsis, sepsis syndrome, trauma/wound, organ transplant,transplant atherosclerosis, transplant rejection, corneal ulcer,chronic/non-healing wound, ulcerative colitis, reperfusion injury(prevent and/or treat), ischemic reperfusion injury (prevent and/ortreat), spinal cord injuries (mitigating effects), cancers,myeloma/multiple myeloma, ovarian cancer, breast cancer, colon cancer,bone cancer osteoarthritis, inflammatory bowel disease, allergicrhinitis, cachexia, diabetes, Alzheimer's disease, implanted prosthesis,biofilm formation, Crohns' disease, dermatitis, acute and chronic,eczema, psoriasis, contact dermatitis, scleroderma, Type I Diabetes,Type II Diabetes, juvenile onset diabetes, prevention of the onset ofdiabetes, diabetic nephropathy, diabetic neuropathy, diabeticretinopathy, erectile dysfunction, macular degeneration, multiplesclerosis, nephropathy, neuropathy, Parkinson's Disease, peripheralvascular disease, and meningitis.

In certain embodiments this invention provides a stent for deliveringdrugs to a vessel in a body. The stent typically comprises a stentframework including a plurality of reservoirs formed therein, and apeptide comprising the amino acid sequence or the retro amino acidsequence of a peptide listed in Tables 2-18 (e.g., Table 4, Table 5, orTable 6) and/or the inverse thereof. In certain embodiments the stentcomprises a a peptide comprising the amino acid sequence of 4F (SEQ IDNO:5) or the inverse thereof. In certain embodiments the active agent iscontained within a polymer. In certain embodiments the stent frameworkcomprises one of a metallic base or a polymeric base. In certainembodiments the stent framework base comprises a material selected fromthe group consisting of stainless steel, nitinol, tantalum, MP35N alloy,platinum, titanium, a suitable biocompatible alloy, a suitablebiocompatible polymer, and a combination thereof. The reservoir(s)comprising said stent can, in some embodiments, comprise micropores(e.g. having a diameter of about 20 microns or less). In certainembodiments the micropores have a diameter in the range of about 20microns to about 50 microns. In various embodiments the micropores havea depth in the range of about 10 to about 50 microns. The micropores, incertain embodiments, extend through the stent framework having anopening on an interior surface of the stent and an opening on anexterior surface of the stent. In various embodiments the stent canfurther comprise a cap layer disposed on the interior surface of thestent framework, the cap layer covering at least a portion of thethrough-holes and providing a barrier characteristic to control anelution rate of a drug in the drug polymer from the interior surface ofthe stent framework. In various embodiments the reservoirs comprisechannels along an exterior surface of the stent framework. In variousembodiments the polymer comprises a first layer of a first drug polymerhaving a first pharmaceutical characteristic and the polymer layercomprises a second drug polymer having a second pharmaceuticalcharacteristic. In certain embodiments the stent further comprises abarrier layer positioned between the polymer comprising the activeagent. In various embodiments a catheter can be coupled to the stentframework. In certain embodiments the catheter can include a balloonused to expand the stent. In certain embodiments the catheter includes asheath that retracts to allow expansion of the stent.

Also provided is a method of manufacturing a drug-polymer stent. Themethod typically involves providing a stent framework; cutting aplurality of reservoirs in the stent framework; applying a compositioncomprising one or more peptides comprising the amino acid sequence orthe retro amino acid sequence of a peptide listed in any of Tables 2-18to at least one reservoir; and drying the composition. The method canfurther involve applying a polymer layer to the dried composition; anddrying the polymer layer.

This invention also provides a method of treating a vascular condition.The method involves positioning a stent as described above, within avessel of a body; expanding the stent; and eluting at least one activeagent (e.g., an active agent from any of Tables 2-18) from at least asurface of the stent.

In certain embodiments, this invention expressly excludes one or more ofthe peptides described in U.S. Pat. Nos. 6,037,323; 4,643,988;6,933,279; 6,930,085; 6,664,230; 3,767,040; 6,037,323; U.S. PatentPublications 2005/0164950; 2004/0266671; 2004/0254120; 2004/0057871;2003/0229015; 2003/0191057; 2003/0171277; 2003/0045460; 2003/0040505;PCT Publications WO 2002/15923; WO 1999/16408; WO 1997/36927; and/or inGarber et al. (1992) Arteriosclerosis and Thrombosis, 12: 886-894, whichare incorporated herein by reference.

DEFINITIONS

The term “treat” when used with reference to treating, e.g. a pathologyor disease refers to the mitigation and/or elimination of one or moresymptoms of that pathology or disease, and/or a reduction in the rate ofonset or severity of one or more symptoms of that pathology or disease,and/or the prevention of that pathology or disease.

The terms “isolated”, “purified”, or “biologically pure” when referringto an isolated polypeptide refer to material that is substantially oressentially free from components that normally accompany it as found inits native state. With respect to nucleic acids and/or polypeptides theterm can refer to nucleic acids or polypeptides that are no longerflanked by the sequences typically flanking them in nature. Chemicallysynthesized polypeptides are “isolated” because they are not found in anative state (e.g. in blood, serum, etc.). In certain embodiments, theterm “isolated” indicates that the polypeptide is not found in nature.

The terms “polypeptide”, “peptide” and “protein” are usedinterchangeably herein to refer to a polymer of amino acid residues. Theterms apply to amino acid polymers in which one or more amino acidresidues is an artificial chemical analogue of a corresponding naturallyoccurring amino acid, as well as to naturally occurring amino acidpolymers.

The term “an amphipathic helical peptide” refers to a peptide comprisingat least one amphipathic helix (amphipathic helical domain). Certainamphipathic helical peptides of this invention can comprise two or more(e.g., 3, 4, 5, etc.) amphipathic helices.

The term “class A amphipathic helix” refers to a protein structure thatforms an α-helix producing a segregation of a polar and nonpolar faceswith the positively charged residues residing at the polar-nonpolarinterface and the negatively charged residues residing at the center ofthe polar face (see, e.g., Segrest et al. (1990) Proteins: Structure,Function, and Genetics 8: 103-117).

“Apolipoprotein J” (apo J) is known by a variety of names includingclusterin, TRPM2, GP80, and SP 40 (see, e.g., Fritz (1995) Pp 112 In:Clusterin: Role in Vertebrate Development, Function, and Adaptation(Harmony JAK Ed.), R. G. Landes, Georgetown, Tex.,). It was firstdescribed as a heterodimeric glycoprotein and a component of thesecreted proteins of cultured rat Sertoli cells (see, e.g., Kissinger etal. (1982) Biol. Reprod.; 27: 233240). The translated product is asingle-chain precursor protein that undergoes intracellular cleavageinto a disulfide-linked 34 kDa α subunit and a 47 kDa β subunit (see,e.g., Collard and Griswold (1987) Biochem., 26: 3297-3303). It has beenassociated with cellular injury, lipid transport, apoptosis and it maybe involved in clearance of cellular debris caused by cell injury ordeath. Clusterin has been shown to bind to a variety of molecules withhigh affinity including lipids, peptides, and proteins and thehydrophobic probe 1-anilino-8-naphthalenesulfonate (Bailey et al. (2001)Biochem., 40: 11828-11840).

The class G amphipathic helix is found in globular proteins, and thus,the name class G. The feature of this class of amphipathic helix is thatit possesses a random distribution of positively charged and negativelycharged residues on the polar face with a narrow nonpolar face. Becauseof the narrow nonpolar face this class does not readily associate withphospholipid (see, e.g., Segrest et al. (1990) Proteins: Structure,Function, and Genetics. 8: 103-117; Erratum (1991) Proteins: Structure,Function and Genetics, 9: 79). Several exchangeable apolipoproteinspossess similar but not identical characteristics to the G amphipathichelix. Similar to the class G amphipathic helix, this other classpossesses a random distribution of positively and negatively chargedresidues on the polar face. However, in contrast to the class Gamphipathic helix which has a narrow nonpolar face, this class has awide nonpolar face that allows this class to readily bind phospholipidand the class is termed G* to differentiate it from the G class ofamphipathic helix (see, e.g., Segrest et al. (1992) J. Lipid Res., 33:141-166; Anantharamaiah et al. (1993) Pp. 109-142 In: The AmphipathicHelix, Epand, R. M. Ed CRC Press, Boca Raton, Fla.). Computer programsto identify and classify amphipathic helical domains have been describedby Jones et al. (1992) J. Lipid Res. 33: 287-296) and include, but arenot limited to the helical wheel program (WHEEL or WHEEL/SNORKEL),helical net program (HELNET, HELNET/SNORKEL, HELNET/Angle), program foraddition of helical wheels (COMBO or COMBO/SNORKEL), program foraddition of helical nets (COMNET, COMNET/SNORKEL, COMBO/SELECT,COMBO/NET), consensus wheel program (CONSENSUS, CONSENSUS/SNORKEL), andthe like.

The term “ameliorating” when used with respect to “ameliorating one ormore symptoms of atherosclerosis” refers to a reduction, prevention, orelimination of one or more symptoms characteristic of atherosclerosisand/or associated pathologies. Such a reduction includes, but is notlimited to a reduction or elimination of oxidized phospholipids, areduction in atherosclerotic plaque formation and rupture, a reductionin clinical events such as heart attack, angina, or stroke, a decreasein hypertension, a decrease in inflammatory protein biosynthesis,reduction in plasma cholesterol, and the like.

The term “enantiomeric amino acids” refers to amino acids that can existin at least two forms that are nonsuperimposable mirror images of eachother. Most amino acids (except glycine) are enantiomeric and exist in aso-called L-form (L amino acid) or D-form (D amino acid). Most naturallyoccurring amino acids are “L” amino acids. The terms “D amino acid” and“L amino acid” are used to refer to absolute configuration of the aminoacid, rather than a particular direction of rotation of plane-polarizedlight. The usage herein is consistent with standard usage by those ofskill in the art. Amino acids are designated herein using standard1-letter or three-letter codes, e.g. as designated in Standard ST.25 inthe Handbook On Industrial Property Information and Documentation.

The term “protecting group” refers to a chemical group that, whenattached to a functional group in an amino acid (e.g. a side chain, analpha amino group, an alpha carboxyl group, etc.) blocks or masks theproperties of that functional group. Preferred amino-terminal protectinggroups include, but are not limited to acetyl, or amino groups. Otheramino-terminal protecting groups include, but are not limited to alkylchains as in fatty acids, propeonyl, formyl and others. Preferredcarboxyl terminal protecting groups include, but are not limited togroups that form amides or esters.

The phrase “protect a phospholipid from oxidation by an oxidizing agent”refers to the ability of a compound to reduce the rate of oxidation of aphospholipid (or the amount of oxidized phospholipid produced) when thatphospholipid is contacted with an oxidizing agent (e.g. hydrogenperoxide, 13-(S)-HPODE, 15-(S)-HPETE, HPODE, HPETE, HODE, HETE, etc.).

The terms “low density lipoprotein” or “LDL” is defined in accordancewith common usage of those of skill in the art. Generally, LDL refers tothe lipid-protein complex which when isolated by ultracentrifugation isfound in the density range d=1.019 to d=1.063.

The terms “high density lipoprotein” or “HDL” is defined in accordancewith common usage of those of skill in the art. Generally “HDL” refersto a lipid-protein complex which when isolated by ultracentrifugation isfound in the density range of d=1.063 to d=1.21.

The term “Group I HDL” refers to a high density lipoprotein orcomponents thereof (e.g. apo A-I, paraoxonase, platelet activatingfactor acetylhydrolase, etc.) that reduce oxidized lipids (e.g. in lowdensity lipoproteins) or that protect oxidized lipids from oxidation byoxidizing agents.

The term “Group II HDL” refers to an HDL that offers reduced activity orno activity in protecting lipids from oxidation or in repairing (e.g.reducing) oxidized lipids.

The term “HDL component” refers to a component (e.g. molecules) thatcomprises a high density lipoprotein (HDL). Assays for HDL that protectlipids from oxidation or that repair (e.g. reduce oxidized lipids) alsoinclude assays for components of HDL (e.g. apo A-I, paraoxonase,platelet activating factor acetylhydrolase, etc.) that display suchactivity.

The term “human apo A-I peptide” refers to a full-length human apo A-Ipeptide or to a fragment or domain thereof comprising a class Aamphipathic helix.

A “monocytic reaction” as used herein refers to monocyte activitycharacteristic of the “inflammatory response” associated withatherosclerotic plaque formation. The monocytic reaction ischaracterized by monocyte adhesion to cells of the vascular wall (e.g.cells of the vascular endothelium), and/or chemotaxis into thesubendothelial space, and/or differentiation of monocytes intomacrophages.

The term “absence of change” when referring to the amount of oxidizedphospholipid refers to the lack of a detectable change, more preferablythe lack of a statistically significant change (e.g. at least at the85%, preferably at least at the 90%, more preferably at least at the95%, and most preferably at least at the 98% or 99% confidence level).The absence of a detectable change can also refer to assays in whichoxidized phospholipid level changes, but not as much as in the absenceof the protein(s) described herein or with reference to other positiveor negative controls.

The following abbreviations may be used herein: PAPC:L-α-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; POVPC:1-palmitoyl-2-(5-oxovaleryl)-sn-glycero-3-phosphocholine; PGPC:1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine; PEIPC:1-palmitoyl-2-(5,6-epoxyisoprostane E₂)-sn-glycero-3-phosphocholine;ChC18:2: cholesteryl linoleate; ChC18:2-OOH: cholesteryl linoleatehydroperoxide; DMPC: 1,2-ditetradecanoyl-rac-glycerol-3-phosphocholine;PON: paraoxonase; HPF: Standardized high power field; PAPC:L-α-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; BL/6:C57BL/6J; C3H:C3H/HeJ.

The term “conservative substitution” is used in reference to proteins orpeptides to reflect amino acid substitutions that do not substantiallyalter the activity (specificity (e.g. for lipoproteins)) or bindingaffinity (e.g. for lipids or lipoproteins)) of the molecule. Typicallyconservative amino acid substitutions involve substitution one aminoacid for another amino acid with similar chemical properties (e.g.charge or hydrophobicity). The following six groups each contain aminoacids that are typical conservative substitutions for one another: 1)Alanine (A), Serine (S), Threonine (T); 2) Aspartic acid (D), Glutamicacid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K);5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and 6)Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

The terms “identical” or percent “identity,” in the context of two ormore nucleic acids or polypeptide sequences, refer to two or moresequences or subsequences that are the same or have a specifiedpercentage of amino acid residues or nucleotides that are the same, whencompared and aligned for maximum correspondence, as measured using oneof the following sequence comparison algorithms or by visual inspection.With respect to the peptides of this invention sequence identity isdetermined over the full length of the peptide.

For sequence comparison, typically one sequence acts as a referencesequence, to which test sequences are compared. When using a sequencecomparison algorithm, test and reference sequences are input into acomputer, subsequence coordinates are designated, if necessary, andsequence algorithm program parameters are designated. The sequencecomparison algorithm then calculates the percent sequence identity forthe test sequence(s) relative to the reference sequence, based on thedesignated program parameters.

Optimal alignment of sequences for comparison can be conducted, e.g., bythe local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482(1981), by the homology alignment algorithm of Needleman & Wunsch, J.Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson& Lipman (1988) Proc. Natl. Acad. Sci. USA 85:2444, by computerizedimplementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA inthe Wisconsin Genetics Software Package, Genetics Computer Group, 575Science Dr., Madison, Wis.), or by visual inspection (see generallyAusubel et al., supra).

One example of a useful algorithm is PILEUP. PILEUP creates a multiplesequence alignment from a group of related sequences using progressive,pairwise alignments to show relationship and percent sequence identity.It also plots a tree or dendogram showing the clustering relationshipsused to create the alignment. PILEUP uses a simplification of theprogressive alignment method of Feng & Doolittle (1987) J. Mol. Evol.35:351-360. The method used is similar to the method described byHiggins & Sharp (1989) CABIOS 5: 151-153. The program can align up to300 sequences, each of a maximum length of 5,000 nucleotides or aminoacids. The multiple alignment procedure begins with the pairwisealignment of the two most similar sequences, producing a cluster of twoaligned sequences. This cluster is then aligned to the next most relatedsequence or cluster of aligned sequences. Two clusters of sequences arealigned by a simple extension of the pairwise alignment of twoindividual sequences. The final alignment is achieved by a series ofprogressive, pairwise alignments. The program is run by designatingspecific sequences and their amino acid or nucleotide coordinates forregions of sequence comparison and by designating the programparameters. For example, a reference sequence can be compared to othertest sequences to determine the percent sequence identity relationshipusing the following parameters: default gap weight (3.00), default gaplength weight (0.10), and weighted end gaps.

Another example of algorithm that is suitable for determining percentsequence identity and sequence similarity is the BLAST algorithm, whichis described in Altschul et al. (1990) J. Mol. Biol. 215: 403-410.Software for performing BLAST analyses is publicly available through theNational Center for Biotechnology Information(http://www.ncbi.nlm.nih.gov/). This algorithm involves firstidentifying high scoring sequence pairs (HSPs) by identifying shortwords of length W in the query sequence, which either match or satisfysome positive-valued threshold score T when aligned with a word of thesame length in a database sequence. T is referred to as the neighborhoodword score threshold (Altschul et al, supra). These initial neighborhoodword hits act as seeds for initiating searches to find longer HSPscontaining them. The word hits are then extended in both directionsalong each sequence for as far as the cumulative alignment score can beincreased. Cumulative scores are calculated using, for nucleotidesequences, the parameters M (reward score for a pair of matchingresidues; always >0) and N (penalty score for mismatching residues;always <0). For amino acid sequences, a scoring matrix is used tocalculate the cumulative score. Extension of the word hits in eachdirection are halted when: the cumulative alignment score falls off bythe quantity X from its maximum achieved value; the cumulative scoregoes to zero or below, due to the accumulation of one or morenegative-scoring residue alignments; or the end of either sequence isreached. The BLAST algorithm parameters W, T, and X determine thesensitivity and speed of the alignment. The BLASTN program (fornucleotide sequences) uses as defaults a wordlength (W) of 11, anexpectation (E) of 10, M=5, N=−4, and a comparison of both strands. Foramino acid sequences, the BLASTP program uses as defaults a wordlength(W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (seeHenikoff & Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915).

In addition to calculating percent sequence identity, the BLASTalgorithm also performs a statistical analysis of the similarity betweentwo sequences (see, e.g., Karlin & Altschul (1993) Proc. Natl. Acad.Sci. USA, 90: 5873-5787). One measure of similarity provided by theBLAST algorithm is the smallest sum probability (P(N)), which providesan indication of the probability by which a match between two nucleotideor amino acid sequences would occur by chance. For example, a nucleicacid is considered similar to a reference sequence if the smallest sumprobability in a comparison of the test nucleic acid to the referencenucleic acid is less than about 0.1, more preferably less than about0.01, and most preferably less than about 0.001.

The phrase “in conjunction with” when used in reference to the use ofone or more drugs in conjunction with one or more active agentsdescribed herein indicates that the drug(s) and the active agent(s) areadministered so that there is at least some chronological overlap intheir physiological activity on the organism. Thus the drug(s) andactive agent(s) can be administered simultaneously and/or sequentially.In sequential administration there may even be some substantial delay(e.g., minutes or even hours or days) before administration of thesecond moiety as long as the first administered drug/agent has exertedsome physiological alteration on the organism when the secondadministered agent is administered or becomes active in the organism.

The phrases “adjacent to each other in a helical wheel diagram of apeptide” or “contiguous in a helical wheel diagram of a peptide” whenreferring to residues in a helical peptide indicates that in the helicalwheel representation the residues appear adjacent or contiguous eventhough they may not be adjacent or contiguous in the linear peptide.Thus, for example, the residues “A, E, K, W, K, and F” are contiguous inthe helical wheel diagrams shown in FIG. 15 even though these residuesare not contiguous in the linear peptide.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a comparison of the effect of D4F (Navab, et al. (2002)Circulation, 105: 290-292) and apo-J peptide 336 made from D amino acids(D-J336*) on the prevention of LDL-induced monocyte chemotactic activityin vitro in a co-incubation experiment. The data are mean ±SD of thenumber of migrated monocytes in nine high power fields in quadruplecultures. (D-J336=Ac-LLEQLNEQFNWVSRLANLTQGE-NH₂, SEQ ID NO:1).

FIG. 2 illustrates the prevention of LDL-induced monocyte chemotacticactivity by pre-treatment of artery wall cells with D-J336 as comparedto D-4F. The data are mean ±SD of the number of migrated monocytes innine high power fields in quadruple cultures.

FIG. 3 illustrates he effect of apo J peptide mimetics on HDL protectivecapacity in LDL receptor null mice. The values are the mean ±SD of thenumber of migrated monocytes in 9 high power fields from each ofquadruple assay wells.

FIG. 4 illustrates protection against LDL-induced monocyte chemotacticactivity by HDL from apo E null mice given oral peptides. The values arethe mean ±SD of the number of migrated monocytes in 9 high power fieldsfrom each of quadruple assay wells. Asterisks indicate significantdifference (p<0.05) as compared to No Peptide mHDL.

FIG. 5 illustrates the effect of oral apo A-I peptide mimetic and apoJpeptide on LDL susceptibility to oxidation. The values are the mean ±SDof the number of migrated monocytes in 9 high power fields from each ofquadruple assay wells. Asterisks indicate significant difference(p<0.05) as compared to No Peptide LDL.

FIG. 6 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on HDL protective capacity. The values are the mean ±SD of thenumber of migrated monocytes in 9 high power fields from each ofquadruple assay wells. Asterisks indicate significant difference(p<0.05) as compared to No Peptide mHDL.

FIG. 7 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on plasma paraoxonase activity. The values are the mean ±SD ofreadings from quadruple plasma aliquots. Asterisks indicate significantdifferences (p<0.05) as compared to No Peptide control plasma.

FIG. 8 shows the effect of oral G* peptides on HDL protective capacityin apoE−/− mice. The values are the mean ±SD of readings from quadrupleplasma aliquots. Asterisks indicate significant differences (p<0.05) ascompared to no peptide control plasma.

FIG. 9 shows the effect of Oral G* peptide, 146-156, on HDL protectivecapacity in ApoE−/− mice.

FIGS. 10A through 10C illustrate helical wheel diagrams of certainpeptides of this invention. FIG. 10A: V²W³A⁵F^(10,17)-D-4F; FIG. 10B:W³-D-4F; FIG. 10C: V²W³F¹⁰-D-4F:

FIG. 11 A standard human LDL (LDL) was added to human artery wallcocultures without (No Addition) or with human HDL (+Control HDL) orwith mouse HDL from apoE null mice given Chow overnight (+Chow HDL), orgiven D-4F in the chow overnight (+D4F HDL) or given G5-D-4F in the chowovernight (+G5 HDL), or given G5,10-D-4F in the chow overnight (+5-10HDL), or given G5,11-D-4F in the chow overnight (+5-11 HDL) and theresulting monocyte chemotactic activity determined as previouslydescribed (Navab et al. (2002) Circulation, 105: 290-292).

FIG. 12 shows that peptides of this invention are effective inmitigating symptoms of diabetes (e.g., blood glucose). Obese Zucker rats26 weeks of age were bled and then treated with daily intraperitonealinjections of D-4F (5.0 mg/kg/day). After 10 days the rats were bledagain plasma glucose and lipid hydroperoxides (LOOH) were determined.*p=0.027; **p=0.0017.

FIG. 13. Sixteen week old Obese Zucker Rats were injected with D-4F (5mg/kg/daily) for 1 week at which time they underwent balloon injury ofthe common carotid artery. Two weeks later the rats were sacrificed andthe intimal media ratio determined.

FIG. 14 demonstrates that the product of the solution phase synthesisscheme is very biologically active in producing HDL and pre-beta HDLthat inhibit LDL-induced monocyte chemotaxis in apo E null mice. ApoEnull mice were fed 5 micrograms of the D-4F synthesized as describedabove (Frgmnt) or the mice were given the same amount of mouse chowwithout D-4F (Chow). Twelve hours after the feeding was started, themice were bled and their plasma was fractionated on FPLC. LDL (100micrograms LDL-cholesterol) was added to cocultures of human artery wallcells alone (LDL) or with a control human HDL (Control HDL) or with HDL(50 micrograms HDL-cholesterol) or post-HDL (pHDL; prebeta HDL) frommice that did (Frgmnt) or did not (Chow) receive the D-4F and themonocyte chemotactic activity produced was determined

FIG. 15 illustrates a helical wheel representation of 4F and reverse(retro) 4F. Reverse-4F is a mirror image of 4F with the relativepositions of the amino acids to each other and to the hydrophilic andhydrophobic faces being identical.

FIG. 16 shows a comparison of the HDL inflammatory index of D-4F versusreverse D-4F.

DETAILED DESCRIPTION

I. Methods of Treatment.

The active agents (e.g. peptides, small organic molecules, amino acidpairs, etc.) described herein are effective for mitigating one or moresymptoms and/or reducing the rate of onset and/or severity of one ormore indications described herein. In particular, the active agents(e.g. peptides, small organic molecules, amino acid pairs, etc.)described herein are effective for mitigating one or more symptoms ofatherosclerosis. Without being bound to a particular theory, it isbelieved that the peptides bind the “seeding molecules” required for theformation of pro-inflammatory oxidized phospholipids such as Ox-PAPC,POVPC, PGPC, and PEIPC.

In addition, since many inflammatory conditions and/or other pathologiesare mediated at least in part by oxidized lipids, we believe that thepeptides of this invention are effective in ameliorating conditions thatare characterized by the formation of biologically active oxidizedlipids. In addition, there are a number of other conditions for whichthe active agents described herein appear to be efficacious.

A number of pathologies for which the active agents described hereinappear to be a palliative and/or a preventative are described below.

A) Atherosclerosis and Associated Pathologies.

We discovered that normal HDL inhibits three steps in the formation ofmildly oxidized LDL. In particular, we demonstrated that treating humanLDL in vitro with apo A-I or an apo A-I mimetic peptide (37pA) removedseeding molecules from the LDL that included HPODE and HPETE. Theseseeding molecules were required for cocultures of human artery wallcells to be able to oxidize LDL and for the LDL to induce the arterywall cells to produce monocyte chemotactic activity. We alsodemonstrated that after injection of apo A-I into mice or infusion intohumans, the LDL isolated from the mice or human volunteers afterinjection/infusion of apo A-I was resistant to oxidation by human arterywall cells and did not induce monocyte chemotactic activity in theartery wall cell cocultures.

The protective function of various active agents of this invention isillustrated in the parent applications (Ser. No. 09/645,454, filed Aug.24, 2000, Ser. No. 09/896,841, filed Jun. 29, 2001, and WO 02/15923(PCT/US01/26497), filed Jun. 29, 2001, see, e.g., FIGS. 1-5 in WO02/15923. FIG. 1, panels A, B, C, and D in WO 02/15923 show theassociation of ¹⁴C-D-5F with blood components in an ApoE null mouse. Itis also demonstrated that HDL from mice that were fed an atherogenicdiet and injected with PBS failed to inhibit the oxidation of human LDLand failed to inhibit LDL-induced monocyte chemotactic activity in humanartery wall coculures. In contrast, HDL from mice fed an atherogenicdiet and injected daily with peptides described herein was as effectivein inhibiting human LDL oxidation and preventing LDL-induced monocytechemotactic activity in the cocultures as was normal human HDL (FIGS. 2Aand 2B in WO 02/15923). In addition, LDL taken from mice fed theatherogenic diet and injected daily with PBS was more readily oxidizedand more readily induced monocyte chemotactic activity than LDL takenfrom mice fed the same diet but injected with 20 μg daily of peptide 5F.The D peptide did not appear to be immunogenic (FIG. 4 in WO 02/15923).

The in vitro responses of human artery wall cells to HDL and LDL frommice fed the atherogenic diet and injected with a peptide according tothis invention are consistent with the protective action shown by suchpeptides in vivo. Despite, similar levels of total cholesterol,LDL-cholesterol, IDL+VLDL-cholesterol, and lower HDL-cholesterol as apercent of total cholesterol, the animals fed the atherogenic diet andinjected with the peptide had significantly lower lesion scores (FIG. 5in WO 02/15923). The peptides of this invention thus preventedprogression of atherosclerotic lesions in mice fed an atherogenic diet.

Thus, in one embodiment, this invention provides methods forameliorating and/or preventing one or more symptoms of atherosclerosisby administering one or more of the active agents described herein.

It is also noted that c-reactive protein, a marker for inflammation, iselevated in congestive heart failure. Also, in congestive heart failurethere is an accumulation of reactive oxygen species and vasomotionabnormalities. Because of their effects in preventing/reducing theformation of various oxidized species and/or because of their effect inimproving vasoreactivity and/or arteriole function (see below) theactive agents described herein will be effective in treating congestiveheart failure.

B) Arteriole/Vascular Indications.

Vessels smaller than even the smallest arteries (i.e., arterioles)thicken, become dysfunctional and cause end organ damage to tissues asdiverse as the brain and the kidney. It is believed the active agentsdescribed herein can function to improve areteriole structure andfunction and/or to slow the rate and/or severity of arterioledysfunction. Without being bound to a particular theory, it is believedthat arteriole dysfunction is a causal factor in various brain andkidney disorders. Use of the agents described herein thus provides amethod to improve the structure and function of arterioles and preservethe function of end organs such as the brain and kidney.

Thus, for example, administration of one or more of the active agentsdescribed herein is expected to reduce one or more symptoms or to slowthe onset or severity of arteriolar disease associated with aging,and/or Alzheimer's disease, and/or Parkinson's disease, and/or withmulti-infarct dementia, and/or subarachnoid hemorrhage, and the like.Similarly, administration of one or more agents described herein isexpected to mitigate one or more symptoms and/or to slow the onsetand/or severity of chronic kidney disease, and/or hypertension.

Similarly, the agents described herein appear to improve vasoreactivity.Because of the improvement of vasoreactivity and/or arteriole function,the agents described herein are suitable for the treatment of peripheralvascular disease, erectile dysfunction, and the like.

C) Pulmonary Indications.

The agents described herein are also suitable for treatment of a varietyof pulmonary indications. These include, but are not limited to chronicobstructive pulmonary disease (COPD), emphysema, pulmonary disease,asthma, idiopathic pulmonary fibrosis, and the like.

D) Mitigation of a Symptom or Condition Associated with CoronaryCalcification and Osteoporosis.

Vascular calcification and osteoporosis often co-exist in the samesubjects (Ouchi et al. (1993) Ann NY Acad. Sci., 676: 297-307; Boukhrisand Becker ('1972) JAMA, 219: 1307-1311; Banks et al. (1994) Eur J ClinInvest., 24: 813-817; Laroche et al. (1994) Clin Rheumatol., 13:611-614; Broulik and Kapitola (1993) Endocr Regul., 27: 57-60; Frye etal. (1992) Bone Mine., 19: 185-194; Barengolts et al. (1998) CalcifTissue Int., 62: 209-213; Burnett and Vasikaran (2002) Ann ClinBiochem., 39: 203-210. Parhami et al. (1997) Arterioscl Thromb VascBiol., 17: 680-687, demonstrated that mildly oxidized LDL (MM-LDL) andthe biologically active lipids in MM-LDL [i.e. oxidized1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine) (Ox-PAPC)],as well as the isoprostane, 8-iso prostaglandin E₂, but not theunoxidized phospholipid (PAPC) or isoprostane 8-iso progstaglandinF_(2α), induced alkaline phosphatase activity and osteoblasticdifferentiation of calcifying vascular cells (CVCs) in vitro, butinhibited the differentiation of MC3T3-E1 bone cells.

The osteon resembles the artery wall in that the osteon is centered onan endothelial cell-lined lumen surrounded by a subendothelial spacecontaining matrix and fibroblast-like cells, which is in turn surroundedby preosteoblasts and osteoblasts occupying a position analogous tosmooth muscle cells in the artery wall (Id.). Trabecular boneosteoblasts also interface with bone marrow subendothelial spaces (Id.).Parhami et al. postulated that lipoproteins could cross the endotheliumof bone arteries and be deposited in the subendothelial space where theycould undergo oxidation as in coronary arteries (Id.). Based on their invitro data they predicted that LDL oxidation in the subendothelial spaceof bone arteries and in bone marrow would lead to reduced osteoblasticdifferentiation and mineralization which would contribute toosteoporosis (Id.). Their hypothesis further predicted that LDL levelswould be positively correlated with osteoporosis as they are withcoronary calcification (Pohle et al. (2001) Circulation, 104:1927-1932), but HDL levels would be negatively correlated withosteoporosis (Parhami et al. (1997) Arterioscl Thromb Vasc Biol., 17:680-687).

In vitro, the osteoblastic differentiation of the marrow stromal cellline M2-10B4 was inhibited by MM-LDL but not native LDL (Parhami et al.(1999) J Bone Miner Res., 14: 2067-2078). When marrow stromal cells fromatherosclerosis susceptible C57BL/6 (BL6) mice fed a low fat chow dietwere cultured there was robust osteogenic differentiation (Id.). Incontrast, when the marrow stromal cells taken from the mice after a highfat, atherogenic diet were cultured they did not undergo osteogenicdifferentiation (Id.). This observation is particularly important sinceit provides a possible explanation for the decreased osteogenicpotential of marrow stromal cells in the development of osteoporosis(Nuttall and Gimble (2000) Bone, 27:177-184). In vivo the decrease inosteogenic potential is accompanied by an increase in adipogenesis inosteoporotic bone (Id.).

It was found that adding D-4F to the drinking water of apoE null micefor 6 weeks dramatically increased trabecular bone mineral density andit is believed that the other active agents of this invention will actsimilarly.

Our data indicate that osteoporosis can be regarded as an“atherosclerosis of bone”. It appears to be a result of the action ofoxidized lipids. HDL destroys these oxidized lipids and promotesosteoblastic differentiation. Our data indicate that administeringactive agent (s) of this invention to a mammal (e.g., in the drinkingwater of apoE null mice) dramatically increases trabecular bone in justa matter of weeks.

This indicates that the active agents, described herein are useful formitigation one or more symptoms of osteoporosis (e.g., for inhibitingdecalcification) or for inducing recalcification of osteoporotic bone.The active agents are also useful as prophylactics to prevent the onsetof symptom(s) of osteoporosis in a mammal (e.g., a patient at risk forosteoporosis).

We believe similar mechanisms are a cause of coronary calcification,e.g., calcific aortic stenosis. Thus, in certain embodiments, thisinvention contemplates the use of the active agents described herein toinhibit or prevent a symptom of a disease such as coronarycalcification, calcific aortic stenosis, osteoporosis, and the like.

E) Inflammatory and Autoimmune Indications.

Chronic inflammatory and/or autoimmune conditions are also characterizedby the formation of a number of reactive oxygen species and are amenableto treatment using one or more of the active agents described herein.Thus, without being bound to a particular theory, we also believe theactive agents described herein are useful, prophylactically ortherapeutically, to mitigate the onset and/or more or more symptoms of avariety of other conditions including, but not limited to rheumatoidarthritis, lupus erythematous, polyarteritis nodosa, polymyalgiarheumatica, scleroderma, multiple sclerosis, and the like.

In certain embodiments, the active agents are useful in mitigating oneor more symptoms caused by, or associated with, an inflammatory responsein these conditions.

Also, in certain embodiments, the active agents are useful in mitigatingone or more symptoms caused by or associated with an inflammatoryresponse associated with AIDS.

F) Infections/Trauma/Transplants.

We have observed that a consequence of influenza infection and otherinfections is the diminution in paraoxonase and platelet activatingacetylhydrolase activity in the HDL. Without being bound by a particulartheory, we believe that, as a result of the loss of these HDL enzymaticactivities and also as a result of the association of pro-oxidantproteins with HDL during the acute phase response, HDL is no longer ableto prevent LDL oxidation and is no longer able to prevent theLDL-induced production of monocyte chemotactic activity by endothelialcells.

We observed that in a subject injected with very low dosages of certainagents of this invention (e.g., 20 micrograms for mice) daily afterinfection with the influenza A virus paraoxonase levels did not fall andthe biologically active oxidized phospholipids were not generated beyondbackground. This indicates that 4F, D4F (and/or other agents of thisinvention) can be administered (e.g. orally or by injection) to patients(including, for example with known coronary artery disease duringinfluenza infection or other events that can generate an acute phaseinflammatory response, e.g. due to viral infection, bacterial infection,trauma, transplant, various autoimmune conditions, etc.) and thus we canprevent by this short term treatment the increased incidence of heartattack and stroke associated with pathologies that generate suchinflammatory states.

In addition, by restoring and/or maintaining paroxonase levels and/ormonocyte activity, the agent(s) of this invention are useful in thetreatment of infection (e.g., viral infection, bacterial infection,fungal infection) and/or the inflammatory pathologies associated withinfection (e.g. meningitis) and/or trauma.

In certain embodiments, because of the combined anti-inflammatoryactivity and anti-infective activity, the agents described herein arealso useful in the treatment of a wound or other trauma, mitigatingadverse effects associated with organ or tissue transplant, and/or organor tissue transplant rejection, and/or implanted prostheses, and/ortransplant atherosclerosis, and/or biofilm formation. In addition, webelieve that L-4F, D-4F, and/or other agents described herein are alsouseful in mitigating the effects of spinal cord injuries.

G) Diabetes and Associated Conditions.

Various active agents described herein have also been observed to showefficacy in reducing and/or preventing one or more symptoms associatedwith diabetes. Thus, in various embodiments, this invention providesmethods of treating (therapeutically and/or prophylactically) diabetesand/or associated pathologies (e.g., Type I diabetes, Type II diabetes,juvenile onset diabetes, diabetic nephropathy, nephropathy, diabeticneuropathy, diabetic retinopathy, and the like.

H) Cancer.

NFκB is a transcription factor that is normally activated in response toproinflammatory cytokines and that regulates the expression of more than200 genes. Many tumor cell lines show constitutive activation of NFκBsignaling. Various studies of mouse models of intestinal, and mammarytumors conclude that activation of the NFκB pathway enhances tumordevelopment and may act primarily in the late stages of tumorigenesis(see, e.g., (2004) Cell 118: 285; (2004) J. Clin. Invest., 114: 569).Inhibition of NFκB signaling suppressed tumor development. Without beingbound to a particular theory, mechanisms for this suppression arebelieved to include an increase in tumor cell apoptosis, reducedexpression of tumor cell growth factors supplied by surrounding stromalcells, and/or abrogation of a tumor cell dedifferentiation program thatis critical for tumor invasion/metastasis.

Without being bound by a particular theory, it is believed theadministration of one or more active agents described herein willinhibit expression and/or secretion, and/or activity of NFκB. Thus, incertain embodiments, this invention provides methods of treating apathology characterized by elevated NFκB by administering one or moreactive agents described herein. Thus, In various embodiments thisinvention contemplates inhibiting NFκB activation associated with cancerby administering one or more active agents described herein, optionallyin combination with appropriate cancer therapeutics.

I) Biochemical Activity.

The active agent(s) described herein have been shown to exhibit a numberof specific biological activities. Thus, for example, they increase hemeoxygenase 1, they increase extracellular superoxide dismutase, theyreduce or prevent the association of myeloperoxidase with apoA-I, theyreduce or prevent the nitrosylation of tyrosine in apoA-I, they renderHDL Anti-inflammatory or more anti-inflammatory, and they increase theformation cycling of pre-β HDL, they promote reverse cholesteroltransport, in particular, reverse cholesterol transport frommacrophages, and they synergize the activity of statins. The activeagents described herein can thus be administered to a mammal to promoteany of these activities, e.g. to treat a condition/pathology whoseseverity, and/or likelihood of onset is reduced by one or more of theseactivities.

J) Mitigation of a Symptom of Atherosclerosis Associated with an AcuteInflammatory Response.

The active agents, of this invention are also useful in a number ofcontexts. For example, we have observed that cardiovascularcomplications (e.g., atherosclerosis, stroke, etc.) frequently accompanyor follow the onset of an acute phase inflammatory response, e.g., suchas that associated with a recurrent inflammatory disease, a viralinfection (e.g., influenza), a bacterial infection, a fungal infection,an organ transplant, a wound or other trauma, and so forth.

Thus, in certain embodiments, this invention contemplates administeringone or more of the active agents described herein to a subject at riskfor, or incurring, an acute inflammatory response and/or at risk for orincurring a symptom of atherosclerosis and/or an associated pathology(e.g., stroke).

Thus, for example, a person having or at risk for coronary disease mayprophylactically be administered a one or more active agents of thisinvention during flu season. A person (or animal) subject to a recurrentinflammatory condition, e.g., rheumatoid arthritis, various autoimmunediseases, etc., can be treated with a one or more agents describedherein to mitigate or prevent the development of atherosclerosis orstroke. A person (or animal) subject to trauma, e.g., acute injury,tissue transplant, etc. can be treated with a polypeptide of thisinvention to mitigate the development of atherosclerosis or stroke.

In certain instances such methods will entail a diagnosis of theoccurrence or risk of an acute inflammatory response. The acuteinflammatory response typically involves alterations in metabolism andgene regulation in the liver. It is a dynamic homeostatic process thatinvolves all of the major systems of the body, in addition to theimmune, cardiovascular and central nervous system. Normally, the acutephase response lasts only a few days; however, in cases of chronic orrecurring inflammation, an aberrant continuation of some aspects of theacute phase response may contribute to the underlying tissue damage thataccompanies the disease, and may also lead to further complications, forexample cardiovascular diseases or protein deposition diseases such asamyloidosis.

An important aspect of the acute phase response is the radically alteredbiosynthetic profile of the liver. Under normal circumstances, the liversynthesizes a characteristic range of plasma proteins at steady stateconcentrations. Many of these proteins have important functions andhigher plasma levels of these acute phase reactants (APRs) or acutephase proteins (APPs) are required during the acute phase responsefollowing an inflammatory stimulus. Although most APRs are synthesizedby hepatocytes, some are produced by other cell types, includingmonocytes, endothelial cells, fibroblasts and adipocytes. Most APRs areinduced between 50% and several-fold over normal levels. In contrast,the major APRs can increase to 1000-fold over normal levels. This groupincludes serum amyloid A (SAA) and either C-reactive protein (CRP) inhumans or its homologue in mice, serum amyloid P component (SAP).So-called negative APRs are decreased in plasma concentration during theacute phase response to allow an increase in the capacity of the liverto synthesize the induced APRs.

In certain embodiments, the acute phase response, or risk therefore isevaluated by measuring one or more APPs. Measuring such markers is wellknown to those of skill in the art, and commercial companies exist thatprovide such measurement (e.g., AGP measured by Cardiotech Services,Louisville, Ky.).

K) Other Indications.

In various embodiments it is contemplated that the active agentsdescribed herein are useful in the treatment (e.g. mitigation and/orprevention) of corneal ulcers, endothelial sloughing, Crohn's disease,acute and chronic dermatitis (including, but not limited to eczemaand/or psoriasis), macular degeneration, neuropathy, scleroderma, andulcerative colitis.

A summary of indications/conditions for which the active agents havebeen shown to be effective and/or are believed to be effective is shownin Table 1. TABLE 1 Summary of conditions in which the active agents(e.g., D-4F) have been shown to be or are believed to be effective.atherosclerosis/symptoms/consequences thereof   plaque formation  lesion formation   myocardial infarction   stroke congestive heartfailure vascular function:   arteriole function   arteriolar disease    associated with aging     associated with alzheimer's disease    associated with chronic kidney disease     associated withhypertension     associated with multi-infarct dementia     associatedwith subarachnoid hemorrhage   peripheral vascular disease pulmonarydisease:   chronic obstructive pulmonary disease (COPD),   emphysema  asthma   idiopathic pulmonary fibrosis   Pulmonary fibrosis   adultrespiratory distress syndrome osteoporosis Paget's disease coronarycalcification autoimmune:     rheumatoid arthritis     polyarteritisnodosa     polymyalgia rheumatica     lupus erythematosus     multiplesclerosis     Wegener's granulomatosis     central nervous systemvasculitis (CNSV)     Sjögren's syndrome     Scleroderma    polymyositis. AIDS inflammatory response infections:   bacterial  fungal   viral   parasitic   influenza     avian flu   viral pneumonia  endotoxic shock syndrome   sepsis   sepsis syndrome   (clinicalsyndrome where it appears that the patient is septic   but no organismsare recovered from the blood) trauma/wound:   organ transplant  transplant atherosclerosis   transplant rejection   corneal ulcer  chronic/non-healing wound   ulcerative colitis   reperfusion injury(prevent and/or treat)   ischemic reperfusion injury (prevent and/ortreat)   spinal cord injuries (mitigating effects) cancers  myeloma/multiple myeloma   ovarian cancer   breast cancer   coloncancer   bone cancer osteoarthritis inflammatory bowel disease allergicrhinitis cachexia diabetes Alzheimer's disease implanted prosthesisbiofilm formation Crohns' disease dermatitis, acute and chronic   eczema  psoriasis   contact dermatitis   scleroderma diabetes and relatedconditions   Type I Diabetes   Type II Diabetes   Juvenile OnsetDiabetes   Prevention of the onset of diabetes   Diabetic Nephropathy  Diabetic Neuropathy   Diabetic Retinopathy erectile dysfunctionmacular degeneration multiple sclerosis nephropathy neuropathyParkinson's Disease peripheral Vascular Disease meningitis Specificbiological activities:   increase Heme Oxygenase 1   increaseextracellular superoxide dismutase   prevent endothelial sloughing  prevent the association of myeloperoxidase with ApoA-I   prevent thenitrosylation of tyrosine in ApoA-I   render HDL anti-inflammatory  improve vasoreactivity   increase the formation of pre-beta HDL  promote reverse cholesterol transport   promote reverse cholesteroltransport from macrophages   synergize the action of statins

It is noted that the conditions listed in Table 1 are intended to beillustrative and not limiting.

L) Administration.

Typically the active agent(s) will be administered to a mammal (e.g., ahuman) in need thereof. Such a mammal will typically include a mammal(e.g. a human) having or at risk for one or more of the pathologiesdescribed herein.

The active agent(s) can be administered, as described herein, accordingto any of a number of standard methods including, but not limited toinjection, suppository, nasal spray, time-release implant, transdermalpatch, and the like. In one particularly preferred embodiment, thepeptide(s) are administered orally (e.g. as a syrup, capsule, ortablet).

The methods involve the administration of a single active agent of thisinvention or the administration of two or more different active agents.The active agents can be provided as monomers (e.g., in separate orcombined formulations), or in dimeric, oligomeric or polymeric forms. Incertain embodiments, the multimeric forms may comprise associatedmonomers (e.g., ionically or hydrophobically linked) while certain othermultimeric forms comprise covalently linked monomers (directly linked orthrough a linker).

While the invention is described with respect to use in humans, it isalso suitable for animal, e.g. veterinary use. Thus certain preferredorganisms include, but are not limited to humans, non-human primates,canines, equines, felines, porcines, ungulates, largomorphs, and thelike.

The methods of this invention are not limited to humans or non-humananimals showing one or more symptom(s) of the pathologies describedherein, but are also useful in a prophylactic context. Thus, the activeagents of this invention can be administered to organisms to prevent theonset/development of one or more symptoms of the pathologies describedherein (e.g., atherosclerosis, stroke, etc.). Particularly preferredsubjects in this context are subjects showing one or more risk factorsfor the pathology. Thus, for example, in the case of atherosclerosisrisk factors include family history, hypertension, obesity, high alcoholconsumption, smoking, high blood cholesterol, high blood triglycerides,elevated blood LDL, VLDL, IDL, or low HDL, diabetes, or a family historyof diabetes, high blood lipids, heart attack, angina or stroke, etc.

II. Active Agents.

A wide variety of active agents are suitable for the treatment of one ormore of the indications discussed above. These agents include, but arenot limited to class A amphipathic helical peptides, class A amphipathichelical peptide mimetics of apoA-I having aromatic or aliphatic residuesin the non-polar face, small peptides including penta-peptides,tetrapeptides, tripeptides, dipeptides and pairs of amino acids, Apo-J(G* peptides), and peptide mimetics, e.g., as described below.

A) Class A Amphipathic Helical Peptides.

In certain embodiments, the activate agents for use in the method ofthis invention include class A amphipathic helical peptides, e.g. asdescribed in U.S. Pat. No. 6,664,230, and PCT Publications WO 02/15923and WO 2004/034977. It was discovered that peptides comprising a class Aamphipathic helix (“class A peptides”), in addition to being capable ofmitigating one or more symptoms of atherosclerosis are also useful inthe treatment of one or more of the other indications described herein.

Class A peptides are characterized by formation of an α-helix thatproduces a segregation of polar and non-polar residues thereby forming apolar and a nonpolar face with the positively charged residues residingat the polar-nonpolar interface and the negatively charged residuesresiding at the center of the polar face (see, e.g., Anantharamaiah(1986) Meth. Enzymol, 128: 626-668). It is noted that the fourth exon ofapo A-I, when folded into 3.667 residues/turn produces a class Aamphipathic helical structure.

One class A peptide, designated 18A (see, e.g., Anantharamaiah (1986)Meth. Enzymol, 128: 626-668) was modified as described herein to producepeptides orally administrable and highly effective at inhibiting orpreventing one or more symptoms of atherosclerosis and/or otherindications described herein. Without being bound by a particulartheory, it is believed that the peptides of this invention may act invivo may by picking up seeding molecule(s) that mitigate oxidation ofLDL.

We determined that increasing the number of Phe residues on thehydrophobic face of 18A would theoretically increase lipid affinity asdetermined by the computation described by Palgunachari et al. (1996)Arteriosclerosis, Thrombosis, & Vascular Biol. 16: 328-338.Theoretically, a systematic substitution of residues in the nonpolarface of 18A with Phe could yield six peptides. Peptides with anadditional 2, 3 and 4 Phe would have theoretical lipid affinity (λ)values of 13, 14 and 15 units, respectively. However, the λ valuesjumped four units if the additional Phe were increased from 4 to 5 (to19 λunits). Increasing to 6 or 7 Phe would produce a less dramaticincrease (to 20 and 21 λunits, respectively).

A number of these class A peptides were made including, the peptidedesignated 4F, D4F, 5F, and D5F, and the like. Various class A peptidesinhibited lesion development in atherosclerosis-susceptible mice. Inaddition, the peptides show varying, but significant degrees of efficacyin mitigating one or more symptoms of the various pathologies describedherein. A number of such peptides are illustrated in Table 2. TABLE 2Illustrative class A amphipathic helical peptides for use in thisinvention. SEQ Peptide ID Name Amino Acid Sequence NO. 18A   D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F 1 2FAc-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 2 3FAc-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 3 3F14Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 4 4FAc-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 5 5FAc-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 6 6FAc-D-W-L-K-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 7 7FAc-D-W-F-K-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 8Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 9Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂ 10Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 11Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 12Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 13Ac-E-W-L-K-L-F-Y-E-K-V-L-E-K-F-K-E-A-F-NH₂ 14Ac-E-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 15Ac-E-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 16Ac-E-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂ 17Ac-E-W-L-K-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 18Ac-E-W-L-K-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 19Ac-E-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 20        AC-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 21        Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 22        Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 23        Ac-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 24        Ac-A-F-Y-D-K-F-F-E-K-F-K-E-F-F-NH₂ 25        Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 26        Ac-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 27        Ac-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂ 28        Ac-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 29        Ac-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 30        Ac-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-NH₂ 31        Ac-L-F-Y-E-K-V-L-E-K-F-K-E-A-F-NH₂ 32        Ac-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 33        Ac-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-NH₂ 34        Ac-A-F-Y-D-K-V-F-E-K-F-K-E-A-F-NH₂ 35        Ac-A-F-Y-D-K-V-F-E-K-L-K-E-F-F-NH₂ 36        Ac-A-F-Y-D-K-V-A-E-K-F-K-E-F-F-NH₂ 37        Ac-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 38Ac-D-W-L-K-A-L-Y-D-K-V-A-E-K-L-K-E-A-L-NH₂ 39Ac-D-W-F-K-A-F-Y-E-K-V-A-E-K-L-K-E-F-F-NH₂ 40Ac-D-W-F-K-A-F-Y-E-K-F-F-E-K-F-K-E-F-F-NH₂ 41Ac-E-W-L-K-A-L-Y-E-K-V-A-E-K-L-K-E-A-L-NH₂ 42Ac-E-W-L-K-A-F-Y-E-K-V-A-E-K-L-K-E-A-F-NH₂ 43Ac-E-W-F-K-A-F-Y-E-K-V-A-E-K-L-K-E-F-F-NH₂ 44Ac-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 45Ac-E-W-L-K-A-F-Y-E-K-F-F-E-K-F-K-E-F-F-NH₂ 46Ac-E-W-F-K-A-F-Y-E-K-F-F-E-K-F-K-E-F-F-NH₂ 47Ac-D-F-L-K-A-W-Y-D-K-V-A-E-K-L-K-E-A-W-NH₂ 48Ac-E-F-L-K-A-W-Y-E-K-V-A-E-K-L-K-E-A-W-NH₂ 49Ac-D-F-W-K-A-W-Y-D-K-V-A-E-K-L-K-E-W-W-NH₂ 50Ac-E-F-W-K-A-W-Y-E-K-V-A-E-K-L-K-E-W-W-NH₂ 51Ac-D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-NH₂ 52Ac-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-NH₂ 53Ac-E-K-L-K-A-F-Y-E-K-V-F-E-W-A-K-E-A-F-NH₂ 54Ac-E-K-W-K-A-V-Y-E-K-F-A-E-A-F-K-E-F-L-NH₂ 55Ac-D-W-L-K-A-F-V-D-K-F-A-E-K-F-K-E-A-Y-NH₂ 56Ac-E-K-W-K-A-V-Y-E-K-F-A-E-A-F-K-E-F-L-NH₂ 57Ac-D-W-L-K-A-F-V-Y-D-K-V-F-K-L-K-E-F-F-NH₂ 58Ac-E-W-L-K-A-F-V-Y-E-K-V-F-K-L-K-E-F-F-NH₂ 59Ac-D-W-L-R-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH₂ 60Ac-E-W-L-R-A-F-Y-E-K-V-A-E-K-L-K-E-A-F-NH₂ 61Ac-D-W-L-K-A-F-Y-D-R-V-A-E-K-L-K-E-A-F-NH₂ 62Ac-E-W-L-K-A-F-Y-E-R-V-A-E-K-L-K-E-A-F-NH₂ 63Ac-D-W-L-K-A-F-Y-D-K-V-A-E-R-L-K-E-A-F-NH₂ 64Ac-E-W-L-K-A-F-Y-E-K-V-A-E-R-L-K-E-A-F-NH₂ 65Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-R-E-A-F-NH₂ 66Ac-E-W-L-K-A-F-Y-E-K-V-A-E-K-L-R-E-A-F-NH₂ 67Ac-D-W-L-K-A-F-Y-D-R-V-A-E-R-L-K-E-A-F-NH₂ 68Ac-E-W-L-K-A-F-Y-E-R-V-A-E-R-L-K-E-A-F-NH₂ 69Ac-D-W-L-R-A-F-Y-D-K-V-A-E-K-L-R-E-A-F-NH₂ 70Ac-E-W-L-R-A-F-Y-E-K-V-A-E-K-L-R-E-A-F-NH₂ 71Ac-D-W-L-R-A-F-Y-D-R-V-A-E-K-L-K-E-A-F-NH₂ 72Ac-E-W-L-R-A-F-Y-E-R-V-A-E-K-L-K-E-A-F-NH₂ 73Ac-D-W-L-K-A-F-Y-D-K-V-A-E-R-L-R-E-A-F-NH₂ 74Ac-E-W-L-K-A-F-Y-E-K-V-A-E-R-L-R-E-A-F-NH₂ 75Ac-D-W-L-R-A-F-Y-D-K-V-A-E-R-L-K-E-A-F-NH₂ 76Ac-E-W-L-R-A-F-Y-E-K-V-A-E-R-L-K-E-A-F-NH₂ 77D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F- P -D-W- 78L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F- P-D-W- 79 L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-FD-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F- P -D-W- 80F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F- P-D-K- 81 L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-FD-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L- P -D-K- 82W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F- P-D-W- 83 F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-FD-W-L-K-A-F-V-Y-D-K-V-F-K-L-K-E-F-F- P -D-W- 84L-K-A-F-V-Y-D-K-V-F-K-L-K-E-F-F D-W-L-K-A-F-Y-D-K-F-A-E-K-F-K-E-F-F- P-D-W- 85 L-K-A-F-Y-D-K-F-A-E-K-F-K-E-F-F Ac-E-W-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-A-F-NH₂ 86 Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-NH₂ 87 Ac-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-NH₂ 88 Ac-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-NH₂ 89NMA-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-NH₂ 90NMA-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-NH₂ 91NMA-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 92NMA-E-W-F-K-A-F-Y-E-K-V-A-E-K-F-K-E-A-F-NH₂ 93NMA-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂ 94NMA-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-NH₂ 95 Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 96NMA-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ Ac-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ 97NMA-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ Ac-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂ 98NMA-A-F-Y-D-K-V-F-E-K-F-K-E-F-F-NH₂  Ac-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂99 NMA-A-F-Y-E-K-V-F-E-K-F-K-E-F-F-NH₂ Ac-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-NH₂ 100NMA-D-W-L-K-A-F-Y-D-K-V-F-E-K-F-NH₂  Ac-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-NH₂101 NMA-E-W-L-K-A-F-Y-E-K-V-F-E-K-F-NH₂ Ac-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-NH₂ 102NMA-L-K-A-F-Y-D-K-V-F-E-K-F-K-E-NH₂  Ac-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-NH₂103 NMA-L-K-A-F-Y-E-K-V-F-E-K-F-K-E-NH₂¹Linkers are underlined.NMA is N-Methyl Anthranilyl.

In certain preferred embodiments, the peptides include variations of 4F((SEQ ID NO:5 in Table 2), also known as L-4F, where all residues are Lform amino acids) or D-4F where one or more residues are D form aminoacids). In any of the peptides described herein, the C-terminus, and/orN-terminus, and/or internal residues can be blocked with one or moreblocking groups as described herein.

While various peptides of Table 2, are illustrated with an acetyl groupor an N-methylanthranilyl group protecting the amino terminus and anamide group protecting the carboxyl terminus, any of these protectinggroups may be eliminated and/or substituted with another protectinggroup as described herein. In particularly preferred embodiments, thepeptides comprise one or more D-form amino acids as described herein. Incertain embodiments, every amino acid (e.g., every enantiomeric aminoacid) of the peptides of Table 2 is a D-form amino acid.

It is also noted that Table 2 is not fully inclusive. Using theteachings provided herein, other suitable class A amphipathic helicalpeptides can routinely be produced (e.g., by conservative orsemi-conservative substitutions (e.g., D replaced by E), extensions,deletions, and the like). Thus, for example, one embodiment utilizestruncations of any one or more of peptides shown herein (e.g., peptidesidentified by SEQ ID Nos:2-20 and 39—in Table 2). Thus, for example, SEQID NO:21 illustrates a peptide comprising 14 amino acids from theC-terminus of 18A comprising one or more D amino acids, while SEQ IDNOS:22-38 illustrate other truncations.

Longer peptides are also suitable. Such longer peptides may entirelyform a class A amphipathic helix, or the class A amphipathic helix(helices) can form one or more domains of the peptide. In addition, thisinvention contemplates multimeric versions of the peptides (e.g.,concatamers). Thus, for example, the peptides illustrated herein can becoupled together (directly or through a linker (e.g., a carbon linker,or one or more amino acids) with one or more intervening amino acids).Illustrative polymeric peptides include 18A-Pro-18A and the peptides ofSEQ ID NOs:78-85, in certain embodiments comprising one or more D aminoacids, more preferably with every amino acid a D amino acid as describedherein and/or having one or both termini protected.

It will also be appreciated in addition to the peptide sequencesexpressly illustrated herein, this invention also contemplates retro andretro-inverso forms of each of these peptides. In retro forms, thedirection of the sequence is reversed. In inverse forms, the chiralityof the constituent amino acids is reversed (i.e., L form amino acidsbecome D form amino acids and D form amino acids become L form aminoacids). In the retro-inverso form, both the order and the chirality ofthe amino acids is reversed. Thus, for example, a retro form of the 4Fpeptide (DWFKAFYDKVAEKFKEAF, SEQ ID NO:5), where the amino terminus isat the aspartate (D) and the carboxyl terminus is at the phenylalanine(F), has the same sequence, but the amino terminus is at thephenylalanine and the carboxy terminus is at the aspartate (i.e.,FAEKFKEAVKDYFAKFWD, SEQ ID NO: 104). Where the 4F peptide comprises allL amino acids, the retro-inverso form will have the sequence shown above(SEQ ID NO:104) and comprise all D form amino acids. As illustrated inthe helical wheel diagrams of FIG. 15, 4F and retroinverso (Rev-4F) aremirror images of each other with identical segregation of the polar andnonpolar faces with the positively charged residues residing at thepolar-nonpolar interface and the negatively charged residues residing atthe center of the polar face. These mirror images of the same polymer ofamino acids are identical in terms of the segregation of the polar andnonpolar faces with the positively charged residues residing at thepolar-nonpolar interface and the negatively charged residues residing atthe center of the polar face. For a discussion of retro- andretro-inverso peptides see, e.g., Chorev and Goodman, (1995) TibTech,13: 439-445.

Where reference is made to a sequence and orientation is not expresslyindicated, the sequence can be viewed as representing the amino acidsequence in the amino to carboxyl orientation, the retro form (i.e., theamino acid sequence in the carboxyl to amino orientation), the retroform where L amino acids are replaced with D amino acids or D aminoacids are replaced with L amino acids, and the retro-inverso form whereboth the order is reversed and the amino acid chirality is reversed.

C) Class A Amphipathic Helical Peptide Mimetics of apoA-I HavingAromatic or Aliphatic Residues in the Non-Polar Face.

In certain embodiments, this invention also provides modified class Aamphipathic helix peptides. Certain preferred peptides incorporate oneor more aromatic residues at the center of the nonpolar face, e.g.,3F^(, Cπ) (as present in 4F), or with one or more aliphatic residues atthe center of the nonpolar face, e.g., 3F^(Iπ), see, e.g., Table 3.Without being bound to a particular theory, we believe the centralaromatic residues on the nonpolar face of the peptide 3F^(Cπ), due tothe presence of π electrons at the center of the nonpolar face, allowwater molecules to penetrate near the hydrophobic lipid alkyl chains ofthe peptide-lipid complex, which in turn would enable the entry ofreactive oxygen species (such as lipid hydroperoxides) shielding themfrom the cell surface. Similarly, we also believe the peptides withaliphatic residues at the center of the nonpolar face, e.g., 3F^(Iπ),will act similarly but not quite as effectively as 3F^(Cπ).

Preferred peptides will convert pro-inflammatory HDL toanti-inflammatory HDL or make anti-inflammatory HDL moreanti-inflammatory, and/or decrease LDL-induced monocyte chemotacticactivity generated by artery wall cells equal to or greater than D4F orother peptides shown in Table 2. TABLE 3 Examples of certain preferredpeptides. SEQ ID Name Sequence NO (3F^(Cπ)) Ac-DKWKAVYDKFAEAFKEFL-NH2105 (3F^(Iπ)) Ac-DKLKAFYDKVFEWAKEAF-NH2 106

C) Other Class A and Some Class Y Amphipathic Helical Peptides.

In certain embodiments this invention also contemplates class aamphipathic helical peptides that have an amino acid compositionidentical to one or more of the class a amphipathic helical peptidesdescribed above. Thus, for example, in certain embodiments thisinvention contemplates peptides having an amino acid compositionidentical to 4F. Thus, in certain embodiments, this invention includesactive agents that comprise a peptide that consists of 18 amino acids,where the 18 amino acids consist of 3 alanines (A), 2 aspartates (D), 2glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine (V), 1tryptophan (W), and 1 tyrosine (Y); and where the peptide forms a classA amphipathic helix; and protects a phospholipid against oxidation by anoxidizing agent. In various embodiments, the peptides comprise least one“D” amino acid residue; and in certain embodiments, the peptidescomprise all “D: form amino acid residues. A variety of such peptidesare illustrated in Table 4. Reverse (retro-), inverse, retro-inverso-,and circularly permuted forms of these peptides are also contemplated.TABLE 4 Illustrative 18 amino acid length class A amphipathic helicalpeptides with the amino acid composition 3 alanines (A), 2 aspartates(D), 2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1 valine(V), 1 tryptophan (W), and 1 tyrosine (Y). SEQ ID Name Sequence NO[Switch D-E]-4F analogs 107 [Switch D-E]-1-4F Ac- E WFKAFY E KVA D KFK DAF-NH2 108 [Switch D-E]-2-4F Ac- E WFKAFYDKVADKFK E AF-NH2 109 [SwitchD-E]-3-4F Ac-DWFKAFY E KVA D KFKEAF-NH2 110 [Switch D-E]-4-4F Ac-DWFKAFYE KVAEKFK D AF-NH2 111 [W-2,F-3 positions reversed] 112 4F-2 Ac-D FWKAFYDKVAEKFKEAF-NH₂ 113 [Switch D-E]-1-4F-2 Ac- E FWKAFY E KVA D KFK DAF-NH2 114 [Switch D-E]-2-4F-2 Ac- E FWKAFYDKVADKFK E AF-NH2 115 [SwitchD-E]-3-4F-2 Ac-DFWKAFY E KVA D KFKEAF-NH2 116 [Switch D-E]-4-4F-2Ac-DFWKAFY E KVAEKFK D AF-NH2 117 [F-6 and Y-7 positions switched] 1184F-3 Ac-DWFKA YF DKVAEKFKEAF-NH₂ 119 [Switch D-E]-1-4F-5 Ac- E WFKAYF EKVA D KFK D AF-NH2 120 [Switch D-E]-2-4F-5 Ac- E WFKAYFDKVADKFK E AF-NH2121 [Switch D-E]-3-4F-5 Ac-DWFKAYF E KVA D KFKEAF-NH2 122 [SwitchD-E]-4-4F-5 Ac-DWFKAYF E KVAEKFK D AF-NH2 123 [Y-7 and 10V positionsswitched] 124 4F-4 Ac-DWFKAF V DK Y AEKFKEAF-NH₂ 125 [Switch D-E]-1-4F-4Ac- E WFKAFV E KYA D KFK D AF-NH2 126 [Switch D-E]-2-4F-4 Ac- EWFKAFVDKYADKFK E AF-NH2 127 [Switch D-E]-3-4F-4 Ac-DWFKAFV E KYA DKFKEAF-NH2 128 [Switch D-E]-4-4F Ac-DWFKAFV E KYAEKFK D AF-NH2 129 [V-10and A-11 switched] 130 4-F-5 Ac-DWFKAFYDK AV EKFKEAF-NH₂ 131 [SwitchD-E]-1-4F-5 Ac- E WFKAFY E KAV D KFK D AF-NH2 132 [Switch D-E]-2-4F-5Ac- E WFKAFYDKAVDKFK E AF-NH2 133 [Switch D-E]-3-4F-5 Ac-DWFKAFY E KAV DKFKEAF-NH2 134 [Switch D-E]-4-4F-5 Ac-DWFKAFY E KAVEKFK D AF-NH2 135[A-11 and F-14 switched] 136 4F-6 Ac-DWFKAFYDKV F EK A KEAF-NH₂ 137[Switch D-E]-1-4F-6 Ac- E WFKAFY E KVF D KAK D AF-NH2 138 [SwitchD-E]-2-4F-6 Ac- E WFKAFYDKVFDKAK E AF-NH2 139 [Switch D-E]-3-4F-6Ac-DWFKAFY E KVF D KAKEAF-NH2 140 [Switch D-E]-4-4F-6 Ac-DWFKAFY EKVFEKAK D AF-NH2 141 [F-14 and A-17 switched] 142 4F-7 Ac-DWFKAFYDKVAEKA KE F F-NH₂ 143 [Switch D-E]-1-4F-7 Ac- E WFKAFY E KVA D KAK D FF-NH2144 [Switch D-E]-2-4F-7 Ac- E WFKAFYDKVADKAK E FF-NH2 145 [SwitchD-E]-3-4F-7 Ac-DWFKAFY E KVA D KAKEFF-NH2 146 [Switch D-E]-4-4F-7Ac-DWFKAFY E KVAEKAK D FF-NH2 147 [A-17 and F-18 switched] 148 4F-8Ac-DWFKAFYDKVAEKFKE FA -NH₂ 149 [Switch D-E]-1-4F-8 Ac- E WFKAFY E KVA DKFK D FA-NH2 150 [Switch D-E]-2-4F-8 Ac- E WFKAFYDKVADKFK E FA-NH2 151[Switch D-E]-3-4F-8 Ac-DWFKAFY E KVA D KFKEFA-NH2 152 [SwitchD-E]-4-4F-8 Ac-DWFKAFY E KVAEKFK D FA-NH2 153 [W-2 and A-17 switched]154 4F-9 Ac-D A FKAFYDKVAEKFKE W F-NH₂ 155 [Switch D-E]-1-4F-9 Ac- EAFKAFY E KVA D KFK D WF-NH2 156 [Switch D-E]-2-4F-9 Ac- E AFKAFYDKVADKFKE WF-NH2 157 [Switch D-E]-3-4F-9 Ac-DAFKAFY E KVA D KFKEWF-NH2 158[Switch D-E]-4-4F-9 Ac-DAFKAFY E KVAEKFK D WF-NH2 159 [W-2 and A-11switched] 160 4F-10 Ac-D A FKAFYDKV W EKFKEAF-NH₂ 161 [SwitchD-E]-1-4F-10 Ac- E AFKAFY E KVW D KFK D AF-NH2 162 [Switch D-E]-2-4F-10Ac- E AFKAFYDKVWDKFK E AF-NH2 163 [Switch D-E]-3-4F-10 Ac-DAFKAFY E KVWD KFKEAF-NH2 164 [Switch D-E]-4-4F-10 Ac-DAFKAFY E KVWEKFK D AF-NH2 165[W-2 and Y-7 switched] 166 4F-11 Ac-D Y FKAF W DKVAEKFKEAF-NH₂ 167[Switch D-E]-1-4F-11 Ac- E YFKAFW E KVA D KFK D AF-NH2 168 [SwitchD-E]-2-4F-11 Ac- E YFKAFWDKVADKFK E AF-NH2 169 [Switch D-E]-3-4F-11Ac-DYFKAFW E KVA D KFKEAF-NH2 170 [Switch D-E]-4-4F-11 Ac-DYFKAFW EKVAEKFK D AF-NH2 171 [F-3 and A-17 switched] 172 4F-12 Ac-DW AKAFYDKVAEKFKE F F-NH₂ 173 [Switch D-E]-1-4F-12 Ac- E WAKAFY E KVA D KFKD FF-NH2 174 [Switch D-E]-2-4F-12 Ac- E WAKAFYDKVADKFK E FF-NH2 175[Switch D-E]-3-4F-12 Ac-DWAKAFY E KVA D KFKEFF-NH2 176 [SwitchD-E]-4-4F-12 Ac-DWAKAFY E KVAEKFK D FF-NH2 177 [F-6 and A-17 switched]178 4F-13 Ac-DWFKA A YDKVAEKFKE F F-NH₂ 179 [Switch D-E]-1-4F-13 Ac- EWFKAAY E KVA D KFK D FF-NH2 180 [Switch D-E]-2-4F-13 Ac- EWFKAAYDKVADKFK E FF-NH2 181 [Switch D-E]-3-4F-13 Ac-DWFKAAY E KVA DKFKEFF-NH2 182 [Switch D-E]-4-4F-13 Ac-DWFKAAY E KVAEKFK D FF-NH2 183[Y-7 and A-17 switched 184 4F-14 Ac-DWFKAF A DKVAEKFKE Y F-NH₂ 185[Switch D-E]-1-4F-14 Ac- E WFKAFA E KVA D KFK D YF-NH2 186 [SwitchD-E]-2-4F-14 Ac- E WFKAFADKVADKFK E YF-NH2 187 Switch D-E]-3-4F-14Ac-DWFKAFA E KVA D KFKEYF-NH2 188 [Switch D-E]-4-4F Ac-DWFKAFA E KVAEKFKD YF-NH2 189 [V-10 and A-17 switched] 190 4F-15 Ac-DWFKAFYDK A AEKFKE VF-NH₂ 191 [Switch D-E]-1-4F-15 Ac- E WFKAFY E KAA D KFK D VF-NH2 192[Switch D-E]-2-4F-15 Ac- E WFKAFYDKAADKFK E VF-NH2 193 [SwitchD-E]-3-4F-15 Ac-DWFKAFY E KAA D KFKEVF-NH2 194 [Switch D-E]-4-4F-15Ac-DWFKAFY E KAAEKFK D VF-NH2 195 [F3 and Y-7 switched] 196 4F-16 Ac-DWY KAF F DKVAEKFKEAF-NH₂ 197 [Switch D-E]-1-4F-16 Ac- E WYKAFF E KVA DKFK D AF-NH2 198 [Switch D-E]-2-4F-16 Ac- E WYKAFFDKVADKFK E AF-NH2 199[Switch D-E]-3-4F-16 Ac-DWYKAFF E KVA D KFKEAF-NH2 200 [SwitchD-E]-4-4F-16 Ac-DWYKAFF E KVAEKFK D AF-NH2 201 [F-3 and V-10 switched]202 4F-17 Ac-DW V KAFYDK F AEKFKEAF-NH₂ 203 [Switch D-E]-1-4F-17 Ac- EWVKAFY E KFA D KFK D AF-NH2 204 [Switch D-E]-2-4F-17 Ac- EWVKAFYDKFADKFK E AF-NH2 205 [Switch D-E]-3-4F-17 Ac-DWVKAFY E KFA DKFKEAF-NH2 206 [Switch D-E]-4-4F-17 Ac-DWVKAFY E KFAEKFK D AF-NH2 207[Y-7 and F-14 switched] 208 4F-18 Ac-DWFKAF F DKVAEK Y KEAF-NH₂ 209[Switch D-E]-1-4F-18 Ac- E WFKAFF E KVA D KYK D AF-NH2 210 [SwitchD-E]-2-4F-18 Ac- E WFKAFFDKVADKYK E AF-NH2 211 [Switch D-E]-3-4F-18Ac-DWFKAFF E KVA D KYKEAF-NH2 212 [Switch D-E]-3-4F-18 Ac-DWFKAFF E KVAD KYKEAF-NH2 213 [Y-7 and F-18 switched] 214 4F-19 Ac-DWFKAF FDKVAEKFKEA Y -NH₂ 215 [Switch D-E]-1-4F-19 Ac- E WFKAFF E KVA D KFK DAY-NH2 216 [Switch D-E]-2-4F-19 Ac- E WFKAFFDKVADKFK E AY-NH2 217[Switch D-E]-3-4F-19 Ac-DWFKAFF E KVA D KFKEAY-NH2 218 [SwitchD-E]-4-4F-19 Ac-DWFKAFF E KVAEKFK D AY-NH2 219 [V-10 and F-18 switched220 4F-20 Ac-DWFKAFYDK F AEKFKEA V -NH₂ 221 [Switch D-E]-1-4F-20 Ac- EWFKAFY E KFA D KFK D AV-NH2 222 [Switch D-E]-2-4F-20 Ac- EWFKAFYDKFADKFK E AV-NH2 223 [Switch D-E]-3-4F-20 Ac-DWFKAFY E KFA DKFKEAV-NH2 224 [Switch D-E]-4-4F-20 Ac-DWFKAFY E KFAEKFK D AV-NH2 225[W-2 and K13 switched] 226 4F-21 Ac-D K FKAFYDKVAEKF W EAF-NH₂ 227[Switch D-E]-1-4F-21 Ac- E KFKAFY E KVA D KFW D AF-NH2 228 [SwitchD-E]-2-4F-21 Ac- E KFKAFYDKVADKFW E AF-NH2 229 [Switch D-E]-3-4F-21Ac-DKFKAFY E KVA D KFWEAF-NH2 230 [Switch D-E]-4-4F-21 Ac-DKFKAFY EKVAEKFW D AF-NH2 231 [W-3, F-13 and K-2 4F] 232 4F-22 Ac-D KWKAFYDKVAEKF F EAF-NH₂ 233 [Switch D-E]-1-4F-22 Ac- E KWKAFY E KVA D KFFD AF-NH2 234 [Switch D-E]-2-4F-22 Ac- E KWKAFYDKVADKFF E AF-NH2 235[Switch D-E]-3-4F-22 Ac-DKWKAFY E KVA D KFFEAF-NH2 236 [SwitchD-E]-4-4F-22 Ac-DKWKAFY E KVAEKFF D AF-NH2 237 [K-2, W10, V-13] 2384F-23 Ac-D K FKAFYDK W AE V FKEAF-NH₂ 239 [Switch D-E]-4F analogs 240[Switch D-E]-1-4F-23 Ac- E KFKAFY E KWA D VFK D AF-NH2 241 [SwitchD-E]-2-4F-23 Ac- E KFKAFYDKWADVFK E AF-NH2 242 [Switch D-E]-3-4F-23Ac-DKFKAFY E KWA D VFKEAF-NH2 243 [Switch D-E]-4-4F-23 Ac-DKFKAFY EKWAEVFK D AF-NH2 244 [K-2, F-13, W-14 4F] 245 4F-24 Ac-D K FKAFYDKVAE FWKEAF-NH 246 [Switch D-E]-4F analogs 247 [Switch D-E]-1-4F-24 Ac- EKFKAFY E KVA D FWK D AF-NH2 248 [Switch D-E]-2-4F-24 Ac- EKFKAFYDKVADFWK E AF-NH2 249 [Switch D-E]-3-4F-24 Ac-DKFKAFY E KVA DFWKEAF-NH2 250 [Switch D-E]-4-4F-24 Ac-DKFKAFY E KVAEFWK D AF-NH2 251Reverse 4F analogs 252 Rev-4F Ac-FAEKFKEAVKDYFAKFWD-NH2 253 [SwitchD-E]-1-Rev-4F Ac-FA D KFK D AVK E YFAKFW E -NH2 254 [SwitchD-E]-2-Rev-4F Ac-FA D KFKEAVKDYFAKFW E -NH2 255 [Switch D-E]-3-Rev-4FAc-FAEKFK D AVK E YFAKFWD-NH2 256 [Switch D-E]-4-Rev-4F Ac-FAEKFK DAVKDYFAKFW E -NH2 257 [A-2 and W-17 switched] 258 Rev-4F-1 Ac-F WEKFKEAVKDYFAKF A D-NH2 259 [Switch D-E]-1-Rev-4F-1 Ac-FW D KFK D AVK EYFAKFA E -NH2 260 [Switch D-E]-2-Rev-4F-1 Ac-FA D KFKEAVKDYFAKFW E -NH2261 [Switch D-E]-3-Rev-4F-1 Ac-FAEKFK D AVK E YFAKFWD-NH2 262 [SwitchD-E]-4-Rev-4F-1 Ac-FAEKFK D AVKDYFAKFW E -NH2 263 [Switch A-2 and F-16]264 Rev-4F-2 Ac-F F EKFKEAVKDYFAK A WD-NH2 265 [Switch D-E]-1-Rev-4F-2Ac-FF D KFK D AVK E YFAKAW E -NH2 266 [Switch D-E]-2-Rev-4F-2 Ac-FF DKFKEAVKDYFAKAW E -NH2 267 [Switch D-E]-3-Rev-4F-2 Ac-FFEKFK D AVK EYFAKAWD-NH2 268 [Switch D-E]-4-Rev-4F-2 Ac-FFEKFK D AVKDYFAKAW E -NH2269 [switch F-5 and A-8] 270 Rev-4F-3 Ac-FAEK A KE F VKDYFAKFWD-NH2 271[Switch D-E]-1-Rev-4F-3 Ac-FA D KAK D FVK E YFAKFW E -NH2 272 [SwitchD-E]-2-Rev-4F-3 Ac-FA D KAKEFVKDYFAKFW E -NH2 273 [SwitchD-E]-3-Rev-4F-3 Ac-FAEKAK D FVK E YFAKFWD-NH2 274 [SwitchD-E]-4-Rev-4F-3 Ac-FAEKAK D FVKDYFAKFW E -NH2 275 [Switch A-8 and V9]276 Rev-4F-4 Ac-FAEKFKE VA KDYFAKFWD-NH2 277 [Switch D-E]-1-Rev-4F-4Ac-FA D KFK D VAK E YFAKFW E -NH2 278 [Switch D-E]-2-Rev-4F-4 Ac-FA DKFKEVAKDYFAKFW E -NH2 279 [Switch D-E]-3-Rev-4F-4 Ac-FAEKFK D VAK EYFAKFWD-NH2 280 [Switch D-E]-4-Rev-4F-4 Ac-FAEKFK D VAKDYFAKFW E -NH2281 [Switch V-9 to Y-12] 282 Rev-4F-5 Ac-FAEKFKEA Y KD V FAKFWD-NH2 283[Switch D-E]-1-Rev-4F-5 Ac-FA D KFK D AYK E VFAKFW E -NH2 284 [SwitchD-E]-2-Rev-4F-5 Ac-FA D KFKEAYKDVFAKFW E -NH2 285 [SwitchD-E]-3-Rev-4F-5 Ac-FAEKFK D AYK E VFAKFWD-NH2 286 [SwitchD-E]-4-Rev-4F-5 Ac-FAEKFK D AYKDVFAKFW E -NH2 287 [Switch Y-12 and F-13]288 Rev-4F-6 Ac-FAEKFKEAVKD FY AKFWD-NH2 289 [Switch D-E]-1-Rev-4F-6Ac-FA D KFK D AVK E FYAKFW E -NH2 290 [Switch D-E]-2-Rev-4F-6 Ac-FA DKFKEAVKDFYAKFW E -NH2 291 [Switch D-E]-3-Rev-4F-6 Ac-FAEKFK D AVK EFYAKFWD-NH2 292 [Switch D-E]-4-Rev-4F-6 Ac-FAEKFK D AVKDFYAKFW E -NH2293 [Switch K-6 and W-17] 294 Rev-4F-7 Ac-FAEKF W EAVKDYFAKF K D-NH2 295[Switch D-E]-1-Rev-4F-7 Ac-FA D KFW D AVK E YFAKFK E -NH2 296 [SwitchD-E]-2-Rev-4F-7 Ac-FA D KFWEAVKDYFAKFK E -NH2 297 [SwitchD-E]-3-Rev-4F-7 Ac-FAEKFW D AVK E YFAKFKD-NH2 298 [SwitchD-E]-4-Rev-4F-7 Ac-FAEKFW D AVKDYFAKFK E -NH2 299 [Switch F-1 and A-2]300 Rev-4F-8 Ac- AF EKFKEAVKDYFAKFWD-NH2 301 [Switch D-E]-1-Rev-4F-8Ac-AF D KFK D AVK E YFAKFW E -NH2 302 [Switch D-E]-2-Rev-4F-8 Ac-AF DKFKEAVKDYFAKFW E -NH2 303 [Switch D-E]-3-Rev-4F-8 Ac-AFEKFK D AVK EYFAKFWD-NH2 304 [Switch D-E]-4-Rev-4F-8 Ac-AFEKFK D AVKDYFAKFW E -NH2305 [F-1 and V-9 are switched] 306 Rev-F-9 Ac- V AEKFKEA F KDYFAKFWD-NH2307 [Switch D-E]-1-Rev-4F-9 Ac-VA D KFK D AFK E YFAKFW E -NH2 308[Switch D-E]-2-Rev-4F-9 Ac-VA D KFKEAFKDYFAKFW E -NH2 309 [SwitchD-E]-3-Rev-4F-9 Ac-VAEKFK D AFK E YFAKFWD-NH2 310 [SwitchD-E]-4-Rev-4F-9 Ac-VAEKFK D AFKDYFAKFW E -NH2 311 [F-1 and Y-12 areswitched] 312 Rev-4F-10 Ac- Y AEKFKEAVKD F FAKFWD-NH2 313 [SwitchD-E]-1-Rev-4F-10 Ac-YA D KFK D AVK E FFAKFW E -NH2 314 [SwitchD-E]-2-Rev-4F-10 Ac-YA D KFKEAVKDFFAKFW E -NH2 315 [SwitchD-E]-3-Rev-4F-10 Ac-YAEKFK D AVK E FFAKFWD-NH2 316 [SwitchD-E]-4-Rev-4F-10 Ac-YAEKFK D AVKDFFAKFW E -NH2 317 [F-1 and A-8 areswitched] 318 Rev-4F-11 Ac- A AEKFKE F VKDYFAKFWD-NH2 319 [SwitchD-E]-1-Rev-4F-11 Ac-AA D KFK D FVK E YFAKFW E -NH2 320 [SwitchD-E]-2-Rev-4F-11 Ac-AA D KFKEFVKDYFAKFW E -NH2 321 [SwitchD-E]-3-Rev-4F-11 Ac-AAEKFK D FVK E YFAKFWD-NH2 322 SwitchD-E]-4-Rev-4F-11 Ac-AAEKFK D FVKDYFAKFW E -NH2 323 [A-2 and F-5 areswitched] 324 Rev-4F-12 Ac-F F EK A KEAVKDYFAKFWD-NH2 325 [SwitchD-E]-1-Rev-4F-12 Ac-FF D KAK D AVK E YFAKFW E -NH2 326 [SwitchD-E]-2-Rev-4F-12 Ac-FF D KAKEAVKDYFAKFW E -NH2 327 [SwitchD-E]-3-Rev-4F-12 Ac-FFEKAK D AVK E YFAKFWD-NH2 328 [SwitchD-E]-4-Rev-4F-12 Ac-FFEKAK D AVKDYFAKFW E -NH2 329 [A-2 and Y12 areswitched 330 Rev-4F-13 Ac-F Y EKFKEAVKD A FAKFWD-NH2 331 [SwitchD-E]-1-Rev-4F-13 Ac-FY D KFK D AVK E AFAKFW E -NH2 332 [SwitchD-E]-2-Rev-4F-13 Ac-FY D KFKEAVKDAFAKFW E -NH2 333 [SwitchD-E]-3-Rev-4F-13 Ac-FYEKFK D AVK E AFAKFWD-NH2 334 [SwitchD-E]-4-Rev-4F-13 Ac-FYEKFK D AVKDAFAKFW E -NH2 335 [A-2 and V-9 areswitched] 336 Rev-4F-14 Ac-F V EKFKEA A KDYFAKFWD-NH2 337 [SwitchD-E]-1-Rev-4F-14 Ac-FV D KFK D AAK E YFAKFW E -NH2 338 [SwitchD-E]-2-Rev-4F-14 Ac-FV D KFKEAAKDYFAKFW E -NH2 339 [SwitchD-E]-3-Rev-4F-14 Ac-FVEKFK D AAK E YFAKFWD-NH2 340 [SwitchD-E]-4-Rev-4F-14 Ac-FVEKFK D AAKDYFAKFW E -NH2 341 [F-5 and Y-12 areswitched] 342 Rev-4F-15 Ac-FAEK Y KEAVKD F FAKFWD-NH2 343 [SwitchD-E]-1-Rev-4F-15 Ac-FA D KYK D AVK E FFAKFW E -NH2 344 [SwitchD-E]-2-Rev-4F-15 Ac-FA D KYKEAVKDFFAKFW E -NH2 345 [SwitchD-E]-3-Rev-4F-15 Ac-FAEKYK D AVK E FFAKFWD-NH2 346 [SwitchD-E]-4-Rev-4F-15 Ac-FAEKYK D AVKDFFAKFW E -NH2 347 [F-5 and V-9 areswitched] 348 Rev-4F-16 Ac-FAEK V KEA F KDYFAKFWD-NH2 349 [SwitchD-E]-1-Rev-4F-16 Ac-FA D KVK D AFK E YFAKFW E -NH2 350 [SwitchD-E]-2-Rev-4F-16 Ac-FA D KVKEAFKDYFAKFW E -NH2 351 [SwitchD-E]-3-Rev-4F-16 Ac-FAEKVK D AFK E YFAKFWD-NH2 352 [SwitchD-E]-4-Rev-4F-16 Ac-FAEKVK D AFKDYFAKFW E -NH2 353 [A-8 and Y-12switched] 354 Rev-4F-17 Ac-FAEKFKE Y VKD A FAKFWD-NH2 355 [SwitchD-E]-1-Rev-4F-17 Ac-FA D KFK D YVK E AFAKFW E -NH2 356 [SwitchD-E]-2-Rev-4F-17 Ac-FA D KFKEYVKDAFAKFW E -NH2 357 [SwitchD-E]-3-Rev-4F-17 Ac-FAEKFK D YVK E AFAKFWD-NH2 358 [SwitchD-E]-4-Rev-4F-17 Ac-FAEKFK D YVKDAFAKFW E -NH2 359 [V-9 and F-13 areswitched] 360 Rev-4F-18 Ac-FAEKFKEA F KDY V AKFWD-NH2 361 [SwitchD-E]-1-Rev-4F-18 Ac-FA D KFK D AFK E YVAKFW E -NH2 362 [SwitchD-E]-2-Rev-4F-18 Ac-FA D KFKEAFKDYVAKFW E -NH2 363 [SwitchD-E]-3-Rev-4F-18 Ac-FAEKFK D AFK E YVAKFWD-NH2 364 [SwitchD-E]-4-Rev-4F-18 Ac-FAEKFK D AFKDYVAKFW E -NH2 365 [V-9 and F-16switched] 366 Rev-4F-19 Ac-FAEKFKEA F KDYFAK V WD-NH2 367 [SwitchD-E]-1-Rev-4F-19 Ac-FA D KFK D AFK E YFAKVW E -NH2 368 [SwitchD-E]-2-Rev-4F-19 Ac-FA D KFKEAFKDYFAKVW E -NH2 369 [SwitchD-E]-3-Rev-4F-19 Ac-FAEKFK D AFK E YFAKVWD-NH2 370 SwitchD-E]-4-Rev-4F-19 Ac-FAEKFK D AFKDYFAKVW E -NH2 371 [Y-12 and F-16 areswitched 372 Rev-4F-20 Ac-FAEKFKEAVKD F FAK Y WD-NH2 373 [SwitchD-E]-1-Rev-4F-20 Ac-FA D KFK D AVK E FFAKYW E -NH2 374 [SwitchD-E]-2-Rev-4F-20 Ac-FA D KFKEAVKDFFAKYW E -NH2 375 [SwitchD-E]-3-Rev-4F-20 Ac-FAEKFK D AVK E FFAKYWD-NH2 376 [SwitchD-E]-4-Rev-4F-20 Ac-FAEKFK D AVKDFFAKYW E -NH2 377 [W-1, F-6 and K-17Rev 4F] 378 Rev-4F-21 Ac- W AEKF F EAVKDYFAKF K D-NH2 379 [SwitchD-E]-1-Rev-4F-7 Ac-WA D KFF D AVK E YFAKFK E -NH2 380 [SwitchD-E]-2-Rev-4F-7 Ac-WA D KFFEAVKDYFAKFK E -NH2 381 [SwitchD-E]-3-Rev-4F-7 Ac-WAEKFF D AVK E YFAKFKD-NH2 382 Switch D-E]-4-Rev-4F-7Ac-WAEKFF D AVKDYFAKFK E -NH2 383 [W-5, F-6 and K-17 Rev-4F] 384Rev-4F-22 Ac-FAEK WF EAVKDYFAKF K D-NH2 385 [Switch D-E]-1-Rev-4F-22Ac-FA D KWF D AVK E YFAKFK E -NH2 386 [Switch D-E]-2-Rev-4F-22 Ac-FA DKWFEAVKDYFAKFK E -NH2 387 [Switch D-E]-3-Rev-4F-22 Ac-FAEKWF D AVK EYFAKFKD-NH2 388 [Switch D-E]-4-Rev-4F-22 Ac-FAEKWF D AVKDYFAKFK E -NH2389 [V-6, W-9, K-17 Rev-4F] 390 Rev-4F-23 Ac-FAEKF V EA W KDYFAKF KD-NH2 391 [Switch D-E]-1-Rev-4F-23 Ac-FA D KFV D AWK E YFAKFK E -NH2 392[Switch D-E]-2-Rev-4F-23 Ac-FA D KFVEAWKDYFAKFK E -NH2 393 [SwitchD-E]-3-Rev-4F-23 Ac-FAEKFV D AWK E YFAKFKD-NH2 394 [SwitchD-E]-4-Rev-4F-23 Ac-FAEKFV D AWKDYFAKFK E -NH2 395 [Y-2, A-4, W-12, K-17Rev-4F] 396 Rev-4F-24 Ac-F Y EKF A EAVKD W FAKF K D-NH2 397 [SwitchD-E]-1-Rev-4F-24 Ac-FY D KFA D AVK E WFAKFK E -NH2 398 [SwitchD-E]-2-Rev-4F-24 Ac-FY D KFAEAVKDWFAKFK E -NH2 399 [SwitchD-E]-3-Rev-4F-24 Ac-FYEKFA D AVK E WFAKFKD-NH2 400 [SwitchD-E]-4-Rev-4F-24 Ac-FYEKFA D AVKDWFAKFK E -NH2 401

Based on the helical wheel diagrams shown in FIG. 15 it is possible toreadily identify biologically active and useful peptides. Thus, forexample, the following peptides have been accurately identified asactive: 3F1; 3F2; 4F the reverse (retro) forms thereof and theretro-inverso forms thereof. Thus, in certain embodiments, thisinvention contemplates active agents comprising a peptide that is 18amino acids in length and forms a class A amphipathic helix where thepeptide has the amino acid composition 2 aspartates, 2 glutamates, 4lysines, 1 tryptophan, 1 tyrosine, no more than one leucine, no morethan 1 valine, no less than 1 and no more than 3 alanines, and with 3 to6 amino acids from the group: phenylalanine, alpha-naphthalanine,beta-naphthalanine, histidine, and contains either 9 or 10 amino acidson the polar face in a helical wheel representation of the class Aamphipathic helix including 4 amino acids with positive charge atneutral pH with two of the positively charged residues residing at theinterface between the polar and non-polar faces and with two of the fourpositively charged residues on the polar face that are contiguous and onthe non-polar face two of the amino acid residues from the group:phenylalanine, alpha-naphthalanine, beta-naphthalanine, histidine arealso contiguous and if there are 4 or more amino acids from this groupon the non-polar face there are also at least 2 residues from this groupthat are not contiguous.

In certain embodiments, this invention also contemplates certain class Yas well as class A amphipathic helical peptides. Class Y amphipathichelical peptides are known to those of skill in the art (see, e.g.,Segrest et al. (1992) J. Lipid Res. 33: 141-166; Oram and Heinecke(2005) Physiol Rev. 85: 1343-1372, and the like). In various embodimentsthese peptides include, but are not limited to an 18 amino acid peptidethat forms a class A amphipathic helix or a class Y amphipathic helixdescribed by formula III (SEQ ID NO:402): DXXKYXXDKXYDKXKDYX III

where the D's are independently Asp or Glu; the Ks are independently Lysor Arg; the Xs are independently Leu, norLeu, Val, Ile, Trp, Phe, Tyr,β-Nal, or α-Nal and all X residues are on the non-polar face (e.g., whenviewed in a helical wheel diagram) except for one that can be on thepolar face between two K residues; the Y's are independently Ala, H is,Ser, Gln, Asn, or Thr non-polar face (e.g., when viewed in a helicalwheel diagram) and the Y's are independently one Ala on the polar face,one H is, one Ser, one Gln one Asn, or one Thr on the polar face (e.g.,when viewed in a helical wheel diagram), where no more than two K are becontiguous (e.g., when viewed in a helical wheel diagram); and where nomore than 3 D's are contiguous (e.g., when viewed in a helical wheeldiagram) and the fourth D is be separated from the other D's by a Y.Illustrative peptides of this kind which include peptides withhistidine, and/or alpha- and/or beta-napthalanine are shown in Table 5.Reverse (retro-), inverse, retro-inverso-, and circularly permuted formsof these peptides are also contemplated. TABLE 5 Illustrates variousclass A and/or class Y peptide analogs with His incorporated into thesequence. SEQ ID Short name Peptide sequence NO [A-5 > H]4F Ac-DWFK HFYDKVAEKFKEAF-NH₂ 403 [A-5 > H, D-E switched]4F Ac- E WFK H FY E KVA DKFK D AF-NH₂ 404 [A-5 > H, D-1 > E]4F Ac- E WFK H FYDKVAEKFKEAF-NH₂ 405[A-5 > H, D-8 > E]4-F Ac-DWFK H FY E KVAEKFKEAF-NH₂ 406 [A-5 > H,E-12 > D]4F Ac-DWFK H FYDKVA D KFKEAF-NH₂ 407 [A-5 > H, E-16 > D]4FAc-DWFK H FYDKVAEKFK D AF-NH₂ 408 [F-3 > H, A-5 > F]-4F Ac-DW H K FFYDKVAEKFKEAF-NH₂ 409 [F-3 > H, A-5 > F, D-E switched]-4F Ac- E W H K FFY E KVA D KFK D AF-NH₂ 410 [F-3 > H, A-5 > F, D-1 > E]-4F Ac- E W H K FFYDKVAEKFKEAF-NH₂ 411 [F-3 > H, A-5 > F, D-8 > E]-4F Ac-DW H K F FY EKVAEKFKEAF-NH₂ 412 [F-3 > H, A-5 > F, E-12 > D]-4F Ac-DW H K F FYDKVA DKFKEAF-NH₂ 413 [F-3 > H, A-5 > F, E-16 > D]-4F Ac-DW H K F FYDKVAEKFK DAF-NH₂ 414 [A-5 > F, F-6 > H]4F Ac-DWFK FH YDKVAEKFKEAF-NH₂ 415[A-5 > F, F-6 > H, D-E switched]4F Ac- E WFK FH Y E KVA D KFK D AF-NH₂416 [[A-5 > F, F-6 > H, D-1 > E]4F Ac- E WFK FH YDKVAEKFKEAF-NH₂ 417[A-5 > F, F-6 > H, D-8 > E]4F Ac-DWFK FH Y E KVAEKFKEAF-NH₂ 418[A-5 > F, F-6 > H, E-12 > D]4F Ac-DWFK FH YDKVA D KFKEAF-NH₂ 419[A-5 > F, F-6 > H, E-16 > D]4F Ac-DWFK FH YDKVAEKFK D AF-NH₂ 420[A-5 > V, V-10 > H]4F Ac-DWFK V FYDK H AEKFKEAF-NH₂ 421 [A-5 > V,V-10 > H, D-E switched]4F Ac- E WFK V FY E K H A D KFK D AF-NH₂ 422[A-5 > V, V-10 > H, D-1 > E]4F Ac- E WFK V FYDK H AEKFKEAF-NH₂ 423[A-5 > V, V-10 > H, D-8 > E]4F Ac-DWFK V FYEK H AEKFKEAF-NH₂ 424[A-5 > V, V-10 > H, E-12 > D]4F Ac-DWFK V FYDK H A D KFKEAF-NH₂ 425[A-5 > V, V-10 > H, E16 > D]4F Ac-DWFK V FYDK H AEKFK D AF-NH₂ 426[[A-17 > H]4F Ac-DWFKAFYDKVAEKFKE H F-NH₂ 427 [A-17 > H, D-E switched]4FAc- E WFKAFY E KVA D KFK DH F-NH₂ 428 [[A-17 > H, D-1 > E]4F Ac- EWFKAFYDKVAEKFKE H F-NH₂ 429 [[A-17 > H, D-8 > E]4F Ac-DWFKAFY E KVAEKFKEH F-NH₂ 430 [[A-17 > H, E-12 > D]4F Ac-DWFKAFYDKVA D KFKE H F-NH₂ 431[[A-17 > H, E16 > D]4F Ac-DWFKAFYDKVAEKFK DH F-NH₂ 432 [A-17 > F,F-18 > H]4F Ac-DWFKAFYDKVAEKFKE FH -NH₂ 433 [A-17 > F, F-18 > H, D-Eswitched]4F Ac- E WFKAFY E KVA D KFK DFH -NH₂ 434 [A-17 > F, F-18 > H,D-1 > E]-4F Ac- E WFKAFYDKVAEKFKE FH -NH₂ 435 [A-17 > F, F-18 > H]4FAc-DWFKAFYDKVAEKFKE FH -NH₂ 436 [A-17 > F, F-18 > H, D-8 > E]-4FAc-DWFKAFY E KVAEKFKE FH -NH₂ 437 [A-17 > F, F-18 > H, E-12 > D]4FAc-DWFKAFYDKVAEKFKE FH -NH₂ 438 [A-17 > F, F-18 > H], E-16 > D]-4FAc-DWFKAFYDKVAEKFK DFH -NH₂ 439 Rev-4F Ac-FAEKFKEAVKDYFAKFWD-NH₂ 440[A-2 > H]Rev4F Ac-F H EKFKEAVKDYFAKFWD-NH₂ 441 Rev-[A-2 > H, D > E]-4FAc-F H EKFKEAVK E YFAKFW E -NH₂ 442 Rev-[A-2 > H, E > D]4F Ac-F HD KFK DAVKDYFAKFWD-NH₂ 443 [A-2 > H, D-E switched]Rev-4F Ac-F HD KFK D AVK EYFAKFW E -NH₂ 444 [A-2 > H, E-3 > D]Rev-4F Ac-F HD KFKEAVKDYFAKFWD-NH₂445 [A-2 > H, E-7 > D]Rev-4F Ac-F H EKFK D AVKDYFAKFWD-NH₂ 446 [A-2 > H,D-11 > E]Rev-4F Ac-F H EKFKEAVK E YFAKFWD-NH₂ 447 [A-2 > H,D-18 > E]Rev-4F Ac-F H EKFKEAVKDYFAKFW E -NH₂ 448 [F-1 > H,A-2 > F]Rev-4F Ac- HF EKFKEAVKDYFAKFWD-NH₂ 449 [F-1 > H, A-2 > F, D-Eswitched]Rev-4F Ac- HFD KFK D AVK E YFAKFW E -NH₂ 450 [F-1 > H, A-2 > F,D > E]Rev-4F Ac- HF EKFKEAVK E YFAKFW E -NH₂ 451 [F-1 > H, A-2 > F,E-3 > D]Rev-4F Ac- HFD KFKEAVKDYFAKFWD-NH₂ 452 [F-1 > H, A-2 > F,E-7 > D]Rev-4F Ac- HF EKFK D AVKDYFAKFWD-NH₂ 453 [F-1 > H, A-2 > F,D-11 > E]Rev-4F Ac- HF EKFKEAVK E YFAKFWD-NH₂ 454 [F-1 > H, A-2 > F,D-18 > E]Rev-4F Ac- HF EKFKEAVKDYFAKFW E -NH₂ 455 [A-2 > F, F-5 > H]RevD-4F Ac-F F EK H KEAVKDYFAKFWD-NH₂ 456 [A-2 > F, F-5 > H, D-Eswitched]RevD-4F Ac-F FD K H K D AVK E YFAKFW E -NH₂ 457 [A-2 > F,F-5 > H, D > E]Rev D-4F Ac-F F EK H KEAVK E YFAKFW E -NH₂ 458 [A-2 > F,F-5 > H, E > D]Rev D-4F Ac-F FD K H K D AVKDYFAKFWD-NH₂ 459 [A-2 > F,F-5 > H, E-3 > D]Rev D-4F Ac-F FD K H KEAVKDYFAKFWD-NH₂ 460 [A-2 > F,F-5 > H, D-11 > E]Rev D-4F Ac-F F EK H K E AVK E YFAKFWD-NH₂ 461[A-2 > F, F-5 > H, D-18 > E]Rev D-4F Ac-F F EK H KEAVKDYFAKFW E -NH₂ 462[A-2 > V, V-9 > H]Rev D-4F Ac-F V EKFKEA H KDYFAKFWD-NH₂ 463 [A-2 > V,V-9 > H, D-E switched]RevD-4F Ac-F VD KFK D A H K E YFAKFW E -NH₂ 464[A-2 > V, V-9 > H, D > E]Rev D-4F Ac-F V EKFKEA H K E YFAKFW E -NH₂ 465[A-2 > V, V-9 > H, E > D]Rev D-4F Ac-F VD KFK D A H KDYFAKFWD-NH₂ 466[A-2 > V, V-9 > H, E-3 > D]Rev D-4F Ac-F VD KFKEA H KDYFAKFWD-NH₂ 467[A-2 > V, V-9 > H, E-7 > D]Rev D-4F Ac-F V EKFK D A H KDYFAKFWD-NH₂ 468[A-2 > V, V-9 > H, D-11 > E]Rev D-4F Ac-F V EKFKEA H KEYFAKFWD-NH₂ 469[A-2 > V, V-9 > H, D-18 > E]Rev D-4F Ac-F V EKFKEA H KDYFAKFW E -NH₂ 470[A-8 > H]Rev-4F Ac-FAEKFKE H VKDYFAKFWD-NH₂ 471 [A-8 > H, D-Eswitched]Rev-4F Ac-FA D KFK DH VKEYFAKFW E -NH₂ 472 [A-8 > H,D > E]Rev-4F Ac-FAEKFKE H VK E YFAKFW E -NH₂ 473 [A-8 > H, E > D]Rev-4FAc-FA D KFK DH VKDYFAKFWD-NH₂ 474 [A-8 > H, E-3 > D]Rev-4F Ac-FA D KFKEH VKDYFAKFWD-NH₂ 475 [A-8 > H, E-7 > D]Rev-4F Ac-FAEKFK DHVKDYFAKFWD-NH₂ 476 [A-8 > H, D-11 > E]Rev-4F Ac-FAEKFKE H VK EYFAKFWD-NH₂ 477 [A-8 > H, D-18 > E]Rev-4F Ac-FAEKFKE H VKDYFAKFW E -NH₂478 [A-8 > F, F-1 3> H]Rev-4F Ac-FAEKFKE F VKDY H AKFWD-NH₂ 479[A-8 > F, F-13 > H, D-E switched]Rev-4F Ac-FA D KFK DF VK E Y H AKFW E-NH₂ 480 [A-8 > F, F-13 > H, E-3 > D]Rev-4F Ac-FA D KFKE F VKDY HAKFWD-NH₂ 481 [A-8 > F, F-13 > H, E-7 > D]Rev-4F Ac-FAEKFK DF VKDY HAKFWD-NH₂ 482 [A-8 > F, F-13 > H, E > D]Rev-4F Ac-FA D KFK DF VKDY HAKFWD-NH₂ 483 [A-8 > F, F-13 > H, D > E]Rev-4F Ac-FAEKFKE F VK E Y HAKFW E -NH₂ 484 [A-8 > F, F-13 > H, D-11 > E]Rev-4F Ac-FAEKFKE F VK E YH AKFWD-NH₂ 485 [A-8 > F, F-13 > H, D-18 > E]Rev-4F Ac-FAEKFKE F VKDY HAKFW E -NH₂ 486 [A-8 > F, F 16 > H]Rev.-4F Ac-FAEKFKE F VKDYFAK H WD-NH₂487 [A-8 > F, F16 > H, D-E switched]Rev.-4F Ac-FA D KFK DF VK E YFAK H WE -NH₂ 488 [A-8 > F, F 16 > H, D > E]Rev.-4F Ac-FAEKFKE F VK E YFAKHW E-NH₂ 489 [A-8 > F, F16 > H, E > D]Rev.-4F Ac-FA D KFK DF VKDYFAK HWD-NH₂ 490 [A-8 > F, F 16 > H, E-3 > D]Rev.-4F Ac-FA D KFKE F VKDYFAK HWD-NH₂ 491 [A-8 > F, F 16 > H, E-7 > D]Rev.-4F Ac-FAEKFK DF VKDYFAK HWD-NH₂ 492 [A-8 > F, F 16 > H, D-11 > E]Rev.-4F Ac-FAEKFKE F VK E YFAK HWD-NH₂ 493 [A-8 > F, F 16 > H, D-18 > E]Rev.-4F Ac-FAEKFKE F VKDYFAK H WE -NH₂ 494 Examples of class A 4F and Rev 4F analogs with beta-Nph.Similarly, alpha-Nph analogs can be designed. Similarly to the aboveanalogs, His can be incorpo- rated to Nph analogs. D > E analogs, E > Danalogs and D-E switch analogs are additional possibilities similarly tothe above described analogs. 4Nph Ac-DW Nph KA Nph YDKVAEK Nph KEA Nph-NH2 495 [D-E switched]4Nph Ac- E W Nph KA Nph Y E KVA D K Nph K D A Nph-NH2 496 [D > E]4Nph Ac- E W Nph KA Nph Y E KVAEK Nph KEA Nph -NH2 497[E > D]4Nph Ac-DW Nph KA Nph YDKVA D K Nph K D A Nph -NH2 498[D-1 > E]4Nph Ac- E W Nph KA Nph YDKVAEK Nph KEA Nph -NH2 499[D-8 > E]4Nph Ac-DW Nph KA Nph Y E KVAEK Nph KEA Nph -NH2 500[E-12 > D]4Nph Ac-DW Nph KA Nph YDKVA D K Nph KEA Nph -NH2 501[E-16 > D]4Nph Ac-DW Nph KA Nph YDKVAEK Nph K D A Nph -NH2 502 Asdescribed above for 4Nph, a minimum of 7 additional analogs for each ofthe analogs given below. [F-3 , 6, > Nph]4F Ac-DW Nph KA NphYDKVAEKFKEAF-NH2 503 [F-14, 18 > Nph]4F Ac-DWFKAFYDKVAEK Nph KEA Nph-NH2 504 [[F-3 > Nph]4F Ac-DW Nph KAFYDKVAEKFKEAF-NH2 505 [F-6 > Nph]4FAc-DWFKA Nph YDKVAEKFKEAF-NH2 506 [F-14 > Nph]4F Ac-DWFKAFYDKVAEK NphKEAF-NH2 507 [F-18 > Nph]4F Ac-DWFKAFYDKVAEKFKEA Nph -NH2 508 For eachof the analog described below, a minimum of 7 additional analogs arepossible as described above by switching D-E, D > E and E > D and singleD or E analogs. Rev-4Nph Ac- Nph AEK Nph KEAVKDY Nph AK Nph WD-NH2 509[F-3, 6 > Nph]Rev 4F Ac- Nph AEK Nph KEAVKDYFAKFWD-NH2 510 [F-13,16]Rev-4F Ac-FAEKFKEAVKDY Nph AK Nph WD-NH2 511 [F-3 > Nph]Rev-4F Ac-Nph AEKFKEAVKDYFAKFWD-NH2 512 [F-6 > Nph]Rev-4F Ac-FAEK NphKEAVKDYFAKFWD-NH2 513 [F-13 > Nph]Rev-4F Ac-FAEKFKEAVKDY Nph AKFWD-NH2514 [F-16 > Nph]Rev-4F Ac-FAEKFKEAVKDYFAK Nph WD-NH2 515 For the analogsdescribed below, additional analogs are possible by incorpo- rating Hisor alpha-Nph and beta-Nph Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E -NH2516 Rev-[E > D]4F Ac-FA D KFK D AVKDYFAKFWD-NH2 517 Rev-R4-4F Ac-FAE RFREAVKDYFAKFWD-NH2 518 Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2 519Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 520 Rev-R14-4F Ac-FAEKFKEAVKDYFAR FWD-NH2 521 Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E -NH2 522Rev-[E > D]4F Ac-FA D KFK D AVKDYFAKFWD-NH2 523 Rev-R4-4F Ac-FAE RFREAVKDYFAKFWD-NH2 524 Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2 525Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 526 Rev-R14-4F Ac-FAEKFKEAVKDYFAR FWD-NH2 527 Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E -NH2 528Rev-[E > D]4F Ac-FA D KFK D AVKDYFAKFWD-NH2 529 Rev-R4-4F Ac-FAE RFREAVKDYFAKFWD-NH2 530 Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2 531Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 532 Rev-R14-4F Ac-FAEKFKEAVKDYFAR FWD-NH2 533 Rev-R4-4F Ac-FAE R FREAVKDYFAKFWD-NH2 534 Rev-R6-4FAc-FAEKF R EAVKDYFAKFWD-NH2 535 Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2536 Rev-R14-4F Ac-FAEKFKEAVKDYFA R FWD-NH2 537 Rev-[D > E]-4FAc-FAEKFKEAVK E YFAKFW E -NH2 538 Rev-[E > D]4F Ac-FA D KFK DAVKDYFAKFWD-NH2 539 Rev-R4-4F Ac-FAE R FREAVKDYFAKFWD-NH2 540 Rev-R6-4FAc-FAEKF R EAVKDYFAKFWD-NH2 541 Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2542 Rev-R14-4F Ac-FAEKFKEAVKDYFA R FWD-NH2 543 For each of the analogsbelow, additional H and Nph analogs are possible using the examplesdescribed above. Each analog can yield 7 analogs with the changesdescribed in the examples given above. Rev3F-2 Ac-LFEKFAEAFKDYVAKWKD-NH2544 RevR4-3F-2 Ac-LFE R FAEAFKDYVAKWKD-NH2 545 RevR10-3F2 Ac-LFEKFAEAF RDYVAKWKD-NH2 546 RevR15-3F-2 Ac-LFEKFAEAFKDYVA R WKD-NH2 547 RevR17-3F-2 Ac-LFEKFAEAFKDYVAKW R D-NH2 548 Rev[D > E]3F2 Ac-LFEKFAEAFK EYVAKWK E -NH2 549 Rev[E > D]3F-2 Ac-LF D KFA D AFKDYVAKWKD-NH2 550Rev-[E3 > D]-3F-2 Ac-LF D KFAEAFKDYVAKWKD-NH2 551 Rev-[E7 > D]-3F-2Ac-LFEKFA D AFKDYVAKWKD-NH2 552 Rev[D11 > E]3F-2 Ac-LFEKFAEAFK EYVAKWKD-NH2 553 Rev-[D18 > E]3F-2 Ac-LFEKFAEAFKDYVAKWK E -NH2 554Rev3F-1 Ac-FAEKAWEFVKDYFAKLKD-NH2 555 RevR4-3F-1 Ac-FAE RAWEFVKDYFAKLKD-NH2 556 RevR10-3F-1 Ac-FAEKAWEFV K DYFAKLKD-NH2 557RevR15-3F-1 Ac-FAEKAWEFVKDYFA K LKD-NH2 558 RevR17-3F-1Ac-FAEKAWEFVKDYFAKL R D-NH2 559 Rev[D > E]3F-1 Ac-FAEKAWEFVK E YFAKLK E-NH2 560 Rev[E > D ]3F-1 Ac-FA D KAW D FVKDYFAKLKD-NH2 561Rev[E3 > D]-3F-1 Ac-FA D KAWEFVKDYFAKLKD-NH2 562 Rev[E7 > D]3F-1Ac-FAEKAW D FVKDYFAKLKD-NH2 563 Rev-[D11 > E]3F-1 Ac-FAEKAWEFVK EYFAKLKD-NH2 564 Rev-[D18 > E]3F-1 Ac-FAEKAWEFVKDYFAKLK E -NH2 565 Rev-5FAc-FFEKFKEFVKDYFAKLWD-NH2 566 Rev-[D > E]5F Ac-FFEKFKEFVK E YFAKLW E-NH2 567 Rev-[E > D]5F Ac-FF D KFK D FVKDYFAKLWD-NH2 568 Rev-R4-5FAc-FFE R FKEFVKDYFAKLWD-NH2 569 Rev-R6-5F Ac-FFEKF R EFVKDYFAKLWD-NH2570 Rev-R10-5F Ac-FFEKFKEFV R DYFAKLWD-NH2 571 Rev-R15-5FAc-FFEKFKEFVKDYFA R LWD-NH2 572 Rev-[E3 > D]-5F Ac-FF DKFKEFVKDYFAKLWD-NH2 573 Rev-[E7 > D]5F Ac-FFEKFK D FVKDYFAKLWD-NH2 574Rev-[D11 > E]-5F Ac-FFEKFKEFVK E YFAKLWD-NH2 575 Rev-[D18 > E]-5FAc-FFEKFKEFVKDYFAKLW E -NH2 576 Rev-5F-2 Ac-F L EKFKEFVKDYFAK F WD-NH2577 Rev-[D > E]-5F-2 Ac-FLEKFKEFVK E YFAKFW E -NH2 578 Rev-[E > D]-5F-2Ac-FL D KFK E FVKDYFAKFWD-NH2 579 Rev-[E3 > D]-5F-2 Ac-FL DKFKEFVKDYFAKFWD-NH2 580 Rev-[E7 > D]-5F-2 Ac-FLEKFK D FVKDYFAKFWD-NH2581 Rev-[D11 > E]-5F-2 Ac-FLEKFKEFVK E YFAKFWD-NH2 582Rev-[D18 > E]-5F-2 Ac-FLEKFKEFVKDYFAKFW E -NH2 583 Rev-R4-5F-2 Ac-FLE RFKEFVKDYFAKFWD-NH2 584 Rev-R6-5F-2 Ac-FLEKF R EFVKDYFAKFWD-NH2 585RevR10-5F-2 Ac-FLEKFKEFV R DYFAKFWD-NH2 586 Rev-R16-5F-2Ac-FLEKFKEFVKDYFA R FWD-NH2 587 Rev-6F Ac-F F EK F KE FF KDYFAKLWD-NH2588 Rev-[D > E]-6F Ac-FFEKFKEFFK E YFAKLW E -NH2 589 Rev-[E > D]-6FAc-FF D KFK D FFKDYFAKLWD-NH2 590 Rev-R4-6F Ac-FFE R FKEFFKDYFAKLWD-NH2591 Rev-R6-6F Ac-FFEKF R EFFKDYFAKLWD-NH2 592 Rev-R10-6F Ac-FFEKFKEFF RDYFAKLWD-NH2 593 Rev-R14-6F Ac-FFERFKEFFKDYFA R LWD-NH2 594Rev-[E3 > D]-6F Ac-FF D KFKEFFKDYFAKLWD-NH2 595 Rev-[E7 > D]-6FAc-FFEKFK D FFKDYFAKLWD-NH2 596 Rev-[D11 > E]-6F Ac-FFEKFKEFFK EYFAKLWD-NH2 597 Rev-[D18 > E]-6F Ac-FFEKFKEFFKDYFAKLW E -NH2 598 Rev-4FAc-FAEKFKEAVKDYFAKFWD-NH2 599 Rev-[D > E]-4F Ac-FAEKFKEAVK E YFAKFW E-NH2 600 Rev-[E > D]4F Ac-FA D KFK D AVKDYFAKFWD-NH2 601 Rev-R4-4FAc-FAE R FREAVKDYFAKFWD-NH2 602 Rev-R6-4F Ac-FAEKF R EAVKDYFAKFWD-NH2603 Rev-R10-4F Ac-FAEKFKEAV R DYFAKFWD-NH2 604 Rev-R14-4FAc-FAEKFKEAVKDYFA R FWD-NH2 605 4F-2 Ac-DKWKAVYDKFAEAFKEFF-NH2 606[D > E]-4F-2 Ac-EKWKAVYEKFAEAFKEFF-NH2 607 [E > D]-4F-2 Ac-DKWKAVYDKFA DAFK D FF-NH2 608 R2-4F-2 Ac-D R WKAVYDKFAEAFKEFF-NH2 609 R4-4F-2 Ac-DKWR AVYDKFAEAFKEFF-NH2 610 R9-4F-2 Ac-DKWKAVYD R FAEAFKEFF-NH2 611R14-4F-2 Ac-DKWKAVYDKFAEAF R EFF-NH2 612 Rev4F-2Ac-FFEKFAEAFKDYVAKWKD-NH2 613 Rev-[D > E]-4F-2 Ac-FFEKFAEAFK E YVAKWK E-NH2 614 Rev-[E > D]-3F-2 Ac-FF D KFA D AFKDYVAKWKD-NH2 615 Rev-R4-4F-2Ac-FFE R FAEAFKDYVAKWKD-NH2 616 Rev-R10-4F-2 Ac-FFERFAEAF R DYVAKWKD-NH2617 Rev-R15-4F-2 Ac-FFEKFAEAFKDYVA R WKD-NH2 618 Rev-R17-4F-2 Ac-FFE RFAEAFKDYVAKW R D-NH2 619 Rev-[E3 > D]-4F-2 Ac-FF D KFAEAFKDYVAKWKD-NH2620 Rev-[E7 > D]-4F-2 Ac-FFEKFA D AFKDYVAKWKD-NH2 621 Rev-[D11 > E]-4F-2Ac-FFERFAEAFK E YVAKWKD-NH2 622 Rev-[D18 > E]-4F-2Ac-FFERFAEAFKDYVAKWKE-NH2 623 Rev-7F Ac-FFEKFKEFFKDYFAKFWD-NH2 624Rev-[E > D]-7F Ac-FF D KFK D FFKDYFAKFWD-NH2 625 Rev-[D > E]-7FAc-FFEKFKEFFK E YFAKFW E -NH2 626 Rev-R4-7F Ac-FFE R FKEFFKDYFAKFWD-NH2627 Rev-R6-7F Ac-FFEKF R EFFKDYFAKFWD-NH2 628 Rev-R10-7F Ac-FFEKFKEFF RDYFAKFWD-NH2 629 Rev-R14-7F Ac-FFEKFKEFFKDYFA R FWD-NH2 630Rev-[E3 > D]-7F Ac-FF D KFKEFFKDYFAKFWD-NH2 631 Rev-[E7 > D]7F Ac-FFEKFKD FFKDYFAKFWD-NH2 632 Rev-[D11 > E]-7F Ac-FFEKFKEFFK E YFAKFWD-NH2 633Rev-[D18 > E]-7F Ac-FFEKFKEFFKDYFAKFW E -NH2 634

It is also noted that any of the peptides described herein can comprisenon-natural amino acids in addition to or instead of the correspondingthe natural amino acids identified herein. Such modifications include,but are not limited to acetylation, amidation, formylation, methylation,sulfation, and the like. Illustrative non-natural amino acids include,but are not limited to Ornithine, norleucine, norvaline, N-methylvaline,6-N-methyllysine, N-methylisoleucine, N-methylglycine, sarcosine,inosine, allo-isoleucine, isodesmolysine, 4-hydroxyproline,3-hydroxyproline, allo-hydroxylysine, hydroxylisine, N-ethylasparagine,N-ethylglycine, 2,3-diaminopropionic acid, 2,2′-diaminopropionic acid,desmosine, 2,4-diaminobutyric acid, 2-aminopimelic acid,3-aminoisobutyric acid, 2-aminoisobutyric acid, 2-aminoheptanoic acid,6-aminocaproic acid, 4-aminobutyric acid, 2-aminobutyric acid,beta-alanine, 3-aminoadipic acid, 2-aminoadipic acid, and the like. Incertain embodiments andy one or more of the “natural” amino acids of thepeptides described herein, can be substituted with the correspondingnon-natural amino acid (e.g. as describe above).

In certain embodiments, this invention contemplates particularly the useof modified lysines. Such modifications include, but are not limited to,biotin modification of epsilon lysines and/or methylation of the epsilonlysines. Illustrative peptide comprising epsilon methylated lysinesinclude, but are not limited to:Ac-D-W-F-K(eCH₃)₂-A-F-Y-D-K(eCH₃)₂-V-A-E-K(eCH₃)₂-F-K(eCH₃)₂-E-A-F-NH(CH₃)₂(SEQ ID NO:635) and:Ac-DWFK(eCH₃)₂AFYDK(eCH₃)₂VAEK(eCH₃)₂FK(eCH₃)₂EAF-NH(CH₃) (SEQ IDNO:636). Other modified amino acids include but are not limited toornithine analogs and homoaminoalanine analogs (instead of (CH₂)₄—NH₂for Lys it can be —(CH₂)₂—NH₂ for Haa and —(CH₂)₃—NH₂ for Orn] and thelike. It is noted that these modifications are illustrative and notintended to be limiting. Illustrative 4F analogues that possess modifiedamino acids are shown in Table 6. TABLE 6 Illustrative 4F analogs thatcomprise modified amino acids. εN-Dimethyl-Lys derivative of 4F(εN-Dime) Ac-D-W-F-K(εN-Dime)-A-F-Y-D-K(εN-Dime)-V-A-E-K 637(εN-Dime)-F-K(εN-Dime)-E-A-F-NH₂Ac-D-W-F-K-(εN-Dime)-A-F-Y-D-K(εN-Dime)-V-A-E-K 638(εN-Dime)-F-K((εN-Dime)-E-A-F-NH-MeAc-D-W-F-K-(εN-Dime)-A-F-Y-D-K(εN-Dime)-V-A-E-K 639(εN-Dime)-F-K(εN-Dime)-E-A-F-N-(Me)₂ εN-Diethyl-Lys derivatives of 4F(εN-Diet) Ac-D-W-F-K(εN-Diet)-A-F-Y-D-K(εN-Diet)-V-A-E-K 640(εN-Diet)-F-K(εN-Diet)-E-A-F-NH₂Ac-D-W-F-K(εN-Diet)-A-F-Y-D-K(εN-Diet)-V-A-E-K 641(εN-Diet)-F-K(εN-Diet)-E-A-F-NH-EtAc-D-W-F-K(εN-Diet)-A-F-Y-D-K(εN-Diet)-V-A-E-K 642(εN-Diet)-F-K(εN-Diet)-E-A-F-NH-(Et)₂ εN-Monomethyl-Lys derivative of 4F(εN-Me) Ac-D-W-F-K(εN-Me)-A-F-Y-D-K(εN-Me)-V-A-E-K(εN- 643Me)-F-K(εN-Me)-E-A-F-NH₂ Ac-D-W-F-K(εN-Me)-A-F-Y-D-K(εN-Me)-V-A-E-K(εN-644 Me)-F-K(εN-Me)-E-A-F-NH-MeAc-D-W-F-K(εN-Me)-A-F-Y-D-K(εN-Me)-V-A-E-K(εN- 645Me)-F-K(εN-Me)-E-A-F-N-(Me)₂ εN-ethylLys derivative of 4F (εN-Et)Ac-D-W-F-K(εN-Et)-A-F-Y-D-K(εN-Et)-V-A-E-K(εN- 646Et)-F-K(εN-Et)-E-A-F-NH₂ Ac-D-W-F-K(εN-Et)-A-F-Y-D-K(εN-Et)-V-A-E-K(εN-647 Et)-F-K(εN-Et)-E-A-F-NH-EtAc-D-W-F-K(εN-Et)-A-F-Y-D-K(εN-Et)-V-A-E-K(εN- 648Et)-F-K(εN-Et)-E-A-F-NH-(Et)₂ HomoLys analogs of 4F (hK) (—CH₂)₅—NH₂Ac-D-W-F-hK-A-F-Y-D-hK-V-A-E-hK-F-hK-E-A-F-NH₂ 649Ac-D-W-F-hK(εN-Dime)-A-F-Y-D-hK(εN-Dime)-V-A-E- 650hK(εN-Dime)-F-hK(εN-Dime)-E-A-F-NH₂Ac-D-W-F-hK(εN-Dime)-A-F-Y-D-hK(εN-Dime)-V-A-E- 651hK(εN-Dime)-F-hK(εN-Dime)-E-A-F-N-(Me)₂Ac-D-W-F-hK(εN-Dime)-A-F-Y-D-hK(εN-Dime)-V-A-E- 652hK(εN-Dime)-F-hK(εN-Dime)-E-A-F-NH-MeAc-D-W-F-hK(εN-Diet)-A-F-Y-D-hK(εN-Diet)-V-A-E- 653hK(εN-Diet)-F-hK(εN-Diet)-E-A-F-NH-EtAc-D-W-F-hK(εN-Me)-A-F-Y-D-hK(εN-Me)-V-A-E-hK 654(εN-Me)-F-hK(εN-Me)-E-A-F-NH₂Ac-D-W-F-hK(εN-Me)-A-F-Y-D-hK(εN-Me)-V-A-E-hK 655(εN-Me)-F-hK(εN-Me)-E-A-F-NH-MeAc-D-W-F-hK(εN-Me)-A-F-Y-D-hK(εN-Me)-V-A-E-hK 656(εN-Me)-F-hK(εN-Me)-E-A-F-N-(Me)₂Ac-D-W-F-hK(εN-Et)-A-F-Y-D-hK(εN-Et)-V-A-E-hK 657(εN-Et)-F-hK(εN-Et)-E-A-F-NH₂Ac-D-W-F-hK(εN-Et)-A-F-Y-D-hK(εN-Et)-V-A-E-hK 658(εN-Et)-F-hK(εN-Et)-E-A-F-NH-EtAc-D-W-F-hK(εN-Et)-A-F-Y-D-hK(εN-Et)-V-A-E-hK 659(εN-Et)-F-hK(εN-Et)-E-A-F-NH-(Et)₂ 4F analogs in which K is replaced O(O = 660 Ornithine, —(CH₂)₃—NH₂)Ac-D-W-F-O-A-F-Y-D-O-V-A-E-O-F-O-E-A-F-NH₂ 661Ac-D-W-F-O(δN-Dime)-A-F-Y-D-O(δN-Dime)-V-A-E-O 662(δN-Dime)-F-O(δN-Dime)-E-A-F-NH₂Ac-D-W-F-O(δN-Dime)-A-F-Y-D-)(δN-Dime)-V-A-E-O 663(δN-Dime)-F-O(δN-Dime)-E-A-F-N-(Me)₂Ac-D-W-F-O(δN-Dime)-A-F-Y-D-O(δN-Dime)-V-A-E-O 664(δN-Dime)-F-O(δN-Dime)-E-A-F-NH-MeAc-D-W-F-O(δN-Diet)-A-F-Y-D-O(δN-Diet)-V-A-E-O 665(δN-Diet)-F-O(δN-Diet)-E-A-F-NH-EtAc-D-W-F-O(δN-Me)-A-F-Y-D-O(δN-Me)-V-A-E-O(δN- 666Me)-F-O(δN-Me)-E-A-F-NH₂ Ac-D-W-F-O(δN-Me)-A-F-Y-D-O(δN-Me)-V-A-E-O(δN-667 Me)-F-O(δN-Me)-E-A-F-NH-MeAc-D-W-F-O(δN-Me)-A-F-Y-D-O(δN-Me)-V-A-E-O(δN- 668Me)-F-O(δN-Me)-E-A-F-N-(Me)₂Ac-D-W-F-O(δN-Et)-A-F-Y-D-O(δN-Et)-V-A-E-O(δN- 669Et)-F-O(δN-Et)-E-A-F-NH₂ Ac-D-W-F-O(δN-Et)-A-F-Y-D-O(δN-Et)-V-A-E-O(δN-670 Et)-F-O(δN-Et)-E-A-F-NH-EtAc-D-W-F-O(δN-Et)-A-F-Y-D-O(δN-Et)-V-A-E-OdεN- 671Et)-F-O(δN-Et)-E-A-F-NH-(Et)₂

The peptides and modifications shown above are intended to beillustrative and not limiting.

D) Smaller Peptides.

It was also a surprising discovery that certain small peptidesconsisting of a minimum of three amino acids preferentially (but notnecessarily) with one or more of the amino acids being theD-stereoisomer of the amino acid, and possessing hydrophobic domains topermit lipid protein interactions, and hydrophilic domains to permit adegree of water solubility also possess significant anti-inflammatoryproperties and are useful in treating one or more of the pathologiesdescribed herein. The “small peptides” typically range in length from 2amino acids to about 15 amino acids, more preferably from about 3 aminoacids to about 10 or 11 amino acids, and most preferably from about 4 toabout 8 or 10 amino acids. In various embodiments the peptides aretypically characterized by having hydrophobic terminal amino acids orterminal amino acids rendered hydrophobic by the attachment of one ormore hydrophobic “protecting” groups. Various “small peptides” aredescribed in copending applications U.S. Ser. No. 10/649,378, filed Aug.26, 2003, and in U.S. Ser. No. 10/913,800, filed on Aug. 6, 2004, and inPCT Application PCT/US2004/026288.

In certain embodiments, the peptides can be characterized by Formula I,below:X¹-X²-X_(n) ³-X⁴  Iwhere, n is 0 or 1, X¹ is a hydrophobic amino acid and/or bears ahydrophobic protecting group, X⁴ is a hydrophobic amino acid and/orbears a hydrophobic protecting group; and when n is 0 X² is an acidic ora basic amino acid; when n is 1: X² and X³ are independently an acidicamino acid, a basic amino acid, an aliphatic amino acid, or an aromaticamino acid such that when X is an acidic amino acid; X³ is a basic aminoacid, an aliphatic amino acid, or an aromatic amino acid; when X² is abasic amino acid; X³ is an acidic amino acid, an aliphatic amino acid,or an aromatic amino acid; and when X² is an aliphatic or aromatic aminoacid, X³ is an acidic amino acid, or a basic amino acid.

Longer peptides (e.g., up to 10, 11, or 15 amino acids) are alsocontemplated within the scope of this invention. Typically where theshorter peptides (e.g., peptides according to formula I) arecharacterized by an acidic, basic, aliphatic, or aromatic amino acid,the longer peptides are characterized by acidic, basic, aliphatic, oraromatic domains comprising two or more amino acids of that type.

1) Functional Properties of Active Small Peptides.

It was a surprising finding of this invention that a number of physicalproperties predict the ability of small peptides (e.g., less than 10amino acids, preferably less than 8 amino acids, more preferably fromabout 3 to about 5 or 6 amino acids) of this invention to render HDLmore anti-inflammatory and to mitigate atherosclerosis and/or otherpathologies characterized by an inflammatory response in a mammal. Thephysical properties include high solubility in ethyl acetate (e.g.,greater than about 4 mg/mL), and solubility in aqueous buffer at pH 7.0.Upon contacting phospholipids such as1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC), in an aqueousenvironment, the particularly effective small peptides induce orparticipate in the formation of particles with a diameter ofapproximately 7.5 nm (±0.1 nm), and/or induce or participate in theformation of stacked bilayers with a bilayer dimension on the order of3.4 to 4.1 nm with spacing between the bilayers in the stack ofapproximately 2 nm, and/or also induce or participate in the formationof vesicular structures of approximately 38 nm). In certain preferredembodiments, the small peptides have a molecular weight of less thanabout 900 Da.

Thus, in certain embodiments, this invention contemplates small peptidesthat ameliorate one or more symptoms of an indication/pathologydescribed herein, e.g., an inflammatory condition, where the peptide(s):ranges in length from about 3 to about 8 amino acids, preferably fromabout 3 to about 6, or 7 amino acids, and more preferably from about 3to about 5 amino acids; are soluble in ethyl acetate at a concentrationgreater than about 4 mg/mL; are soluble in aqueous buffer at pH 7.0;when contacted with a phospholipid in an aqueous environment, formparticles with a diameter of approximately 7.5 nm and/or form stackedbilayers with a bilayer dimension on the order of 3.4 to 4.1 nm withspacing between the bilayers in the stack of approximately 2 nm; have amolecular weight less than about 900 daltons; convert pro-inflammatoryHDL to anti-inflammatory HDL or make anti-inflammatory HDL moreanti-inflammatory; and do not have the amino acid sequenceLys-Arg-Asp-Ser (SEQ ID NO:801), especially in which Lys-Arg-Asp and Serare all L amino acids. In certain embodiments, these small peptidesprotect a phospholipid against oxidation by an oxidizing agent.

While these small peptides need not be so limited, in certainembodiments, these small peptides can include the small peptidesdescribed below.

2) Tripeptides.

It was discovered that certain tripeptides (3 amino acid peptides) canbe synthesized that show desirable properties as described herein (e.g.,the ability to convert pro-inflammatory HDL to anti-inflammatory HDL,the ability to decrease LDL-induced monocyte chemotactic activitygenerated by artery wall cells, the ability to increase pre-beta HDL,etc.). In certain embodiments, the peptides are characterized by formulaI, wherein N is zero, shown below as Formula II:X¹-X²-X⁴  IIwhere the end amino acids (X¹ and X⁴) are hydrophobic either because ofa hydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). In certain embodiments,the X² amino acid is either acidic (e.g., aspartic acid, glutamic acid,etc.) or basic (e.g., histidine, arginine, lysine, etc.). The peptidecan be all L-amino acids or include one or more or all D-amino acids.

Certain preferred tripeptides of this invention include, but are notlimited to the peptides shown in Table 7. TABLE 7 Examples of certainpreferred tripeptides bearing hydrophobic blocking groups and acidic,basic, or histidine central amino acids. SEQ ID X¹ X² X³ X⁴ NOBoc-Lys(εBoc) Arg Ser(tBu)-OtBu 672 Boc-Lys(εBoc) Arg Thr(tBu)-OtBu 673Boc-Trp Arg Ile-OtBu 674 Boc-Trp Arg Leu-OtBu 675 Boc-Phe Arg Ile-OtBu676 Boc-Phe Arg Leu-OtBu 677 Boc-Lys(εBoc) Glu Ser(tBu)-OtBu 678Boc-Lys(εBoc) Glu Thr(tBu)-OtBu 679 Boc-Lys(εBoc) Asp Ser(tBu)-OtBu 680Boc-Lys(εBoc) Asp Thr(tBu)-OtBu 681 Boc-Lys(εBoc) Arg Ser(tBu)-OtBu 682Boc-Lys(εBoc) Arg Thr(tBu)-OtBu 683 Boc-Leu Glu Ser(tBu)-OtBu 684Boc-Leu Glu Thr(tBu)-OtBu 685 Fmoc-Trp Arg Ser(tBu)-OtBu 686 Fmoc-TrpAsp Ser(tBu)-OtBu 687 Fmoc-Trp Glu Ser(tBu)-OtBu 688 Fmoc-Trp ArgSer(tBu)-OtBu 689 Boc-Lys(εBoc) Glu Leu-OtBu 690 Fmoc-Leu ArgSer(tBu)-OtBu 691 Fmoc-Leu Asp Ser(tBu)-OtBu 692 Fmoc-Leu GluSer(tBu)-OtBu 693 Fmoc-Leu Arg Ser(tBu)-OtBu 694 Fmoc-Leu ArgThr(tBu)-OtBu 695 Boc-Glu Asp Tyr(tBu)-OtBu 696 Fmoc-Lys(εFmoc) ArgSer(tBu)-OtBu 697 Fmoc-Trp Arg Ile-OtBu 698 Fmoc-Trp Arg Leu-OtBu 699Fmoc-Phe Arg Ile-OtBu 700 Fmoc-Phe Arg Leu-OtBu 701 Boc-Trp Arg Phe-OtBu702 Boc-Trp Arg Tyr-OtBu 703 Fmoc-Trp Arg Phe-OtBu 704 Fmoc-Trp ArgTyr-OtBu 705 Boc-Orn(δBoc) Arg Ser(tBu)-OtBu 706 Nicotinyl Lys(εBoc) ArgSer(tBu)-OtBu 707 Nicotinyl Lys(εBoc) Arg Thr(tBu)-OtBu 708 Fmoc-Leu AspThr(tBu)-OtBu 709 Fmoc-Leu Glu Thr(tBu)-OtBu 710 Fmoc-Leu ArgThr(tBu)-OtBu 711 Fmoc-norLeu Arg Ser(tBu)-OtBu 712 Fmoc-norLeu AspSer(tBu)-OtBu 713 Fmoc-norLeu Glu Ser(tBu)-OtBu 714 Fmoc-Lys(εBoc) ArgSer(tBu)-OtBu 715 Fmoc-Lys(εBoc) Arg Thr(tBu)-OtBu 716 Fmoc-Lys(εBoc)Glu Ser(tBu)-OtBu 717 Fmoc-Lys(εBoc) Glu Thr(tBu)-OtBu 718Fmoc-Lys(εBoc) Asp Ser(tBu)-OtBu 719 Fmoc-Lys(εBoc) Asp Thr(tBu)-OtBu720 Fmoc-Lys(εBoc) Glu Leu-OtBu 721 Fmoc-Lys(εBoc) Arg Leu-OtBu 722Fmoc-Lys(εFmoc) Arg Thr(tBu)-OtBu 723 Fmoc-Lys(εFmoc) Glu Ser(tBu)-OtBu724 Fmoc-Lys(εFmoc) Glu Thr(tBu)-OtBu 725 Fmoc-Lys(εFmoc) AspSer(tBu)-OtBu 726 Fmoc-Lys(εFmoc) Asp Thr(tBu)-OtBu 727 Fmoc-Lys(εFmoc)Arg Ser(tBu)-OtBu 728 Fmoc-Lys(εFmoc)) Glu Leu-OtBu 729 Boc-Lys(εFmoc)Asp Ser(tBu)-OtBu 730 Boc-Lys(εFmoc) Asp Thr(tBu)-OtBu 731Boc-Lys(εFmoc) Arg Thr(tBu)-OtBu 732 Boc-Lys(εFmoc) Glu Leu-OtBu 733Boc-Orn(δFmoc) Glu Ser(tBu)-OtBu 734 Boc-Orn(δFmoc) Asp Ser(tBu)-OtBu735 Boc-Orn(δFmoc) Asp Thr(tBu)-OtBu 736 Boc-Orn(δFmoc) ArgThr(tBu)-OtBu 737 Boc-Orn(δFmoc) Glu Thr(tBu)-OtBu 738 Fmoc-Trp AspIle-OtBu 739 Fmoc-Trp Arg Ile-OtBu 740 Fmoc-Trp Glu Ile-OtBu 741Fmoc-Trp Asp Leu-OtBu 742 Fmoc-Trp Glu Leu-OtBu 743 Fmoc-Phe AspIle-OtBu 744 Fmoc-Phe Asp Leu-OtBu 745 Fmoc-Phe Glu Leu-OtBu 746Fmoc-Trp Arg Phe-OtBu 747 Fmoc-Trp Glu Phe-OtBu 748 Fmoc-Trp AspPhe-OtBu 749 Fmoc-Trp Asp Tyr-OtBu 750 Fmoc-Trp Arg Tyr-OtBu 751Fmoc-Trp Glu Tyr-OtBu 752 Fmoc-Trp Arg Thr(tBu)-OtBu 753 Fmoc-Trp AspThr(tBu)-OtBu 754 Fmoc-Trp Glu Thr(tBu)-OtBu 755 Boc-Phe Arg norLeu-OtBu756 Boc-Phe Glu norLeu-OtBu 757 Fmoc-Phe Asp norLeu-OtBu 758 Boc-Glu HisTyr(tBu)-OtBu 759 Boc-Leu His Ser(tBu)-OtBu 760 Boc-Leu HisThr(tBu)-OtBu 761 Boc-Lys(εBoc) His Ser(tBu)-OtBu 762 Boc-Lys(εBoc) HisThr(tBu)-OtBu 763 Boc-Lys(εBoc) His Leu-OtBu 764 Boc-Lys(εFmoc) HisSer(tBu)-OtBu 765 Boc-Lys(εFmoc) His Thr(tBu)-OtBu 766 Boc-Lys(εFmoc)His Leu-OtBu 767 Boc-Orn(δBoc) His Ser(tBu)-OtBu 768 Boc-Orn(δFmoc) HisThr(tBu)-OtBu 769 Boc-Phe His Ile-OtBu 770 Boc-Phe His Leu-OtBu 771Boc-Phe His norLeu-OtBu 772 Boc-Phe Lys Leu-OtBu 773 Boc-Trp HisIle-OtBu 774 Boc-Trp His Leu-OtBu 775 Boc-Trp His Phe-OtBu 776 Boc-TrpHis Tyr-OtBu 777 Boc-Phe Lys Leu-OtBu 778 Fmoc-Lys(εFmoc) HisSer(tBu)-OtBu 779 Fmoc-Lys(εFmoc) His Thr(tBu)-OtBu 780 Fmoc-Lys(εFmoc)His Leu-OtBu 781 Fmoc-Leu His Ser(tBu)-OtBu 782 Fmoc-Leu HisThr(tBu)-OtBu 783 Fmoc-Lys(εBoc) His Ser(tBu)-OtBu 784 Fmoc-Lys(εBoc)His Thr(tBu)-OtBu 785 Fmoc-Lys(εBoc) His Leu-OtBu 786 Fmoc-Lys(εFmoc)His Ser(tBu)-OtBu 787 Fmoc-Lys(εFmoc) His Thr(tBu)-OtBu 788 Fmoc-norLeuHis Ser(tBu)-OtBu 789 Fmoc-Phe His Ile-OtBu 790 Fmoc-Phe His Leu-OtBu791 Fmoc-Phe His norLeu-OtBu 792 Fmoc-Trp His Ser(tBu)-OtBu 793 Fmoc-TrpHis Ile-OtBu 794 Fmoc-Trp His Leu-OtBu 795 Fmoc-Trp His Phe-OtBu 796Fmoc-Trp His Tyr-OtBu 797 Fmoc-Trp His Thr(tBu)-OtBu 798 NicotinylLys(εBoc) His Ser(tBu)-OtBu 799 Nicotinyl Lys(εBoc) His Thr(tBu)-OtBu800

While the peptides of Table 7 are illustrated with particular protectinggroups, it is noted that these groups may be substituted with otherprotecting groups as described herein and/or one or more of the shownprotecting group can be eliminated.

3) Small Peptides with Central Acidic and Basic Amino Acids.

In certain embodiments, the peptides of this invention range from fouramino acids to about ten amino acids. The terminal amino acids aretypically hydrophobic either because of a hydrophobic side chain orbecause the terminal amino acids bear one or more hydrophobic protectinggroups end amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, Nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). Typically, the centralportion of the peptide comprises a basic amino acid and an acidic aminoacid (e.g., in a 4 mer) or a basic domain and/or an acidic domain in alonger molecule.

These four-mers can be represented by Formula I in which X¹ and X⁴ arehydrophobic and/or bear hydrophobic protecting group(s) as describedherein and X² is acidic while X³ is basic or X² is basic while X³ isacidic. The peptide can be all L-amino acids or include one or more orall D-amino acids.

Certain preferred of this invention include, but are not limited to thepeptides shown in Table 8. TABLE 8 Illustrative examples of smallpeptides with central acidic and basic amino acids. SEQ ID X1 X2 X3 X4NO Boc-Lys(εBoc) Arg Asp Ser(tBu)-OtBu 801 Boc-Lys(εBoc) Arg AspThr(tBu)-OtBu 802 Boc-Trp Arg Asp Ile-OtBu 803 Boc-Trp Arg Asp Leu-OtBu804 Boc-Phe Arg Asp Leu-OtBu 805 Boc-Phe Arg Asp Ile-OtBu 806 Boc-PheArg Asp norLeu-OtBu 807 Boc-Phe Arg Glu norLeu-OtBu 808 Boc-Phe Arg GluIle-OtBu 809 Boc-Phe Asp Arg Ile-OtBu 810 Boc-Phe Glu Arg Ile-OtBu 811Boc-Phe Asp Arg Leu-OtBu 812 Boc-Phe Arg Glu Leu-OtBu 813 Boc-Phe GluArg Leu-OtBu 814 Boc-Phe Asp Arg norLeu-OtBu 815 Boc-Phe Glu ArgnorLeu-OtBu 816 Boc-Lys(εBoc) Glu Arg Ser(tBu)-OtBu 817 Boc-Lys(εBoc)Glu Arg Thr(tBu)-OtBu 818 Boc-Lys(εBoc) Asp Arg Ser(tBu)-OtBu 819Boc-Lys(εBoc) Asp Arg Thr(tBu)-OtBu 820 Boc-Lys(εBoc) Arg GluSer(tBu)-OtBu 821 Boc-Lys(εBoc) Arg Glu Thr(tBu)-OtBu 822 Boc-Leu GluArg Ser(tBu)-OtBu 823 Boc-Leu Glu Arg Thr(tBu)-OtBu 824 Fmoc-Trp Arg AspSer(tBu)-OtBu 825 Fmoc-Trp Asp Arg Ser(tBu)-OtBu 826 Fmoc-Trp Glu ArgSer(tBu)-OtBu 827 Fmoc-Trp Arg Glu Ser(tBu)-OtBu 828 Boc-Lys(εBoc) GluArg Leu-OtBu 829 Fmoc-Leu Arg Asp Ser(tBu)-OtBu 830 Fmoc-Leu Asp ArgSer(tBu)-OtBu 831 Fmoc-Leu Glu Arg Ser(tBu)-OtBu 832 Fmoc-Leu Arg GluSer(tBu)-OtBu 833 Fmoc-Leu Arg Asp Thr(tBu)-OtBu 834 Boc-Glu Asp ArgTyr(tBu)-OtBu 835 Fmoc-Lys(εFmoc) Arg Asp Ser(tBu)-OtBu 836 Fmoc-Trp ArgAsp Ile-OtBu 837 Fmoc-Trp Arg Asp Leu-OtBu 838 Fmoc-Phe Arg Asp Ile-OtBu839 Fmoc-Phe Arg Asp Leu-OtBu 840 Boc-Trp Arg Asp Phe-OtBu 841 Boc-TrpArg Asp Tyr-OtBu 842 Fmoc-Trp Arg Asp Phe-OtBu 843 Fmoc-Trp Arg AspTyr-OtBu 844 Boc-Orn(δBoc) Arg Glu Ser(tBu)-OtBu 845 Nicotinyl Lys(εBoc)Arg Asp Ser(tBu)-OtBu 846 Nicotinyl Lys(εBoc) Arg Asp Thr(tBu)-OtBu 847Fmoc-Leu Asp Arg Thr(tBu)-OtBu 848 Fmoc-Leu Glu Arg Thr(tBu)-OtBu 849Fmoc-Leu Arg Glu Thr(tBu)-OtBu 850 Fmoc-norLeu Arg Asp Ser(tBu)-OtBu 851Fmoc-norLeu Asp Arg Ser(tBu)-OtBu 852 Fmoc-norLeu Glu Arg Ser(tBu)-OtBu853 Fmoc-norLeu Arg Glu Ser(tBu)-OtBu 854 Fmoc-Lys(εBoc) Arg AspSer(tBu)-OtBu 855 Fmoc-Lys(εBoc) Arg Asp Thr(tBu)-OtBu 856Fmoc-Lys(εBoc) Glu Arg Ser(tBu)-OtBu 857 Fmoc-Lys(εBoc) Glu ArgThr(tBu)-OtBu 858 Fmoc-Lys(εBoc) Asp Arg Ser(tBu)-OtBu 859Fmoc-Lys(εBoc) Asp Arg Thr(tBu)-OtBu 860 Fmoc-Lys(εBoc) Arg GluSer(tBu)-OtBu 861 Fmoc-Lys(εBoc) Arg Glu Thr(tBu)-OtBu 862Fmoc-Lys(εBoc) Glu Arg Leu-OtBu 863 Fmoc-Lys(εBoc) Arg Glu Leu-OtBu 864Fmoc-Lys(εFmoc) Arg Asp Thr(tBu)-OtBu 865 Fmoc-Lys(εFmoc) Glu ArgSer(tBu)-OtBu 866 Fmoc-Lys(εFmoc) Glu Arg Thr(tBu)-OtBu 867Fmoc-Lys(εFmoc) Asp Arg Ser(tBu)-OtBu 868 Fmoc-Lys(εFmoc) Asp ArgThr(tBu)-OtBu 869 Fmoc-Lys(εFmoc) Arg Glu Ser(tBu)-OtBu 870Fmoc-Lys(εFmoc) Arg Glu Thr(tBu)-OtBu 871 Fmoc-Lys(εFmoc)) Glu ArgLeu-OtBu 872 Boc-Lys(εFmoc) Arg Asp Ser(tBu)-OtBu 873 Boc-Lys(εFmoc) ArgAsp Thr(tBu)-OtBu 874 Boc-Lys(εFmoc) Glu Arg Ser(tBu)-OtBu 875Boc-Lys(εFmoc) Glu Arg Thr(tBu)-OtBu 876 Boc-Lys(εFmoc) Asp ArgSer(tBu)-OtBu 877 Boc-Lys(εFmoc) Asp Arg Thr(tBu)-OtBu 878Boc-Lys(εFmoc) Arg Glu Ser(tBu)-OtBu 879 Boc-Lys(εFmoc) Arg GluThr(tBu)-OtBu 880 Boc-Lys(εFmoc) Glu Arg Leu-OtBu 881 Boc-Orn(δFmoc) ArgGlu Ser(tBu)-OtBu 882 Boc-Orn(δFmoc) Glu Arg Ser(tBu)-OtBu 883Boc-Orn(δFmoc) Arg Asp Ser(tBu)-OtBu 884 Boc-Orn(δFmoc) Asp ArgSer(tBu)-OtBu 885 Boc-Orn(δFmoc) Asp Arg Thr(tBu)-OtBu 886Boc-Orn(δFmoc) Arg Asp Thr(tBu)-OtBu 887 Boc-Orn(δFmoc) Glu ArgThr(tBu)-OtBu 888 Boc-Orn(δFmoc) Arg Glu Thr(tBu)-OtBu 889 Fmoc-Trp AspArg Ile-OtBu 890 Fmoc-Trp Arg Glu Ile-OtBu 891 Fmoc-Trp Glu Arg Ile-OtBu892 Fmoc-Trp Asp Arg Leu-OtBu 893 Fmoc-Trp Arg Glu Leu-OtBu 894 Fmoc-TrpGlu Arg Leu-OtBu 895 Fmoc-Phe Asp Arg Ile-OtBu 896 Fmoc-Phe Arg GluIle-OtBu 897 Fmoc-Phe Glu Arg Ile-OtBu 898 Fmoc-Phe Asp Arg Leu-OtBu 899Fmoc-Phe Arg Glu Leu-OtBu 900 Fmoc-Phe Glu Arg Leu-OtBu 901 Fmoc-Trp ArgAsp Phe-OtBu 902 Fmoc-Trp Arg Glu Phe-OtBu 903 Fmoc-Trp Glu Arg Phe-OtBu904 Fmoc-Trp Asp Arg Tyr-OtBu 905 Fmoc-Trp Arg Glu Tyr-OtBu 906 Fmoc-TrpGlu Arg Tyr-OtBu 907 Fmoc-Trp Arg Asp Thr(tBu)-OtBu 908 Fmoc-Trp Asp ArgThr(tBu)-OtBu 909 Fmoc-Trp Arg Glu Thr(tBu)-OtBu 910 Fmoc-Trp Glu ArgThr(tBu)-OtBu 911 Fmoc-Phe Arg Asp norLeu-OtBu 912 Fmoc-Phe Arg GlunorLeu-OtBu 913 Boc-Phe Lys Asp Leu-OtBu 914 Boc-Phe Asp Lys Leu-OtBu915 Boc-Phe Lys Glu Leu-OtBu 916 Boc-Phe Glu Lys Leu-OtBu 917 Boc-PheLys Asp Ile-OtBu 918 Boc-Phe Asp Lys Ile-OtBu 919 Boc-Phe Lys GluIle-OtBu 920 Boc-Phe Glu Lys Ile-OtBu 921 Boc-Phe Lys Asp norLeu-OtBu922 Boc-Phe Asp Lys norLeu-OtBu 923 Boc-Phe Lys Glu norLeu-OtBu 924Boc-Phe Glu Lys norLeu-OtBu 925 Boc-Phe His Asp Leu-OtBu 926 Boc-Phe AspHis Leu-OtBu 927 Boc-Phe His Glu Leu-OtBu 928 Boc-Phe Glu His Leu-OtBu929 Boc-Phe His Asp Ile-OtBu 930 Boc-Phe Asp His Ile-OtBu 931 Boc-PheHis Glu Ile-OtBu 932 Boc-Phe Glu His Ile-OtBu 933 Boc-Phe His AspnorLeu-OtBu 934 Boc-Phe Asp His norLeu-OtBu 935 Boc-Phe His GlunorLeu-OtBu 936 Boc-Phe Glu His norLeu-OtBu 937 Boc-Lys(εBoc) Lys AspSer(tBu)-OtBu 938 Boc-Lys(εBoc) Asp Lys Ser(tBu)-OtBu 939 Boc-Lys(εBoc)Lys Glu Ser(tBu)-OtBu 940 Boc-Lys(εBoc) Glu Lys Ser(tBu)-OtBu 941Boc-Lys(εBoc) His Asp Ser(tBu)-OtBu 942 Boc-Lys(εBoc) Asp HisSer(tBu)-OtBu 943 Boc-Lys(εBoc) His Glu Ser(tBu)-OtBu 944 Boc-Lys(εBoc)Glu His Ser(tBu)-OtBu 945

While the peptides of Table 8 are illustrated with particular protectinggroups, it is noted that these groups may be substituted with otherprotecting groups as described herein and/or one or more of the shownprotecting group can be eliminated.

4) Small Peptides Having Either an Acidic or Basic Amino Acid in theCenter Together with a Central Aliphatic Amino Acid.

In certain embodiments, the peptides of this invention range from fouramino acids to about ten amino acids. The terminal amino acids aretypically hydrophobic either because of a hydrophobic side chain orbecause the terminal amino acids bear one or more hydrophobic protectinggroups. End amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, Nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). Typically, the centralportion of the peptide comprises a basic or acidic amino acid and analiphatic amino acid (e.g., in a 4 mer) or a basic domain or an acidicdomain and an aliphatic domain in a longer molecule.

These four-mers can be represented by Formula I in which X¹ and X⁴ arehydrophobic and/or bear hydrophobic protecting group(s) as describedherein and X² is acidic or basic while X³ is aliphatic or X² isaliphatic while X³ is acidic or basic. The peptide can be all L-aminoacids or include one, or more, or all D-amino acids.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 9. TABLE 9 Examples of certainpreferred peptides having either an acidic or basic amino acid in thecenter together with a central aliphatic amino acid. SEQ ID X¹ X² X³ X⁴NO Fmoc-Lys(εBoc) Leu Arg Ser(tBu)-OtBu 946 Fmoc-Lys(εBoc) Arg LeuSer(tBu)-OtBu 947 Fmoc-Lys(εBoc) Leu Arg Thr(tBu)-OtBu 948Fmoc-Lys(εBoc) Arg Leu Thr(tBu)-OtBu 949 Fmoc-Lys(εBoc) Glu LeuSer(tBu)-OtBu 950 Fmoc-Lys(εBoc) Leu Glu Ser(tBu)-OtBu 951Fmoc-Lys(εBoc) Glu Leu Thr(tBu)-OtBu 952 Fmoc-Lys(εBoc) Leu GluThr(tBu)-OtBu 953 Fmoc-Lys(εFmoc) Leu Arg Ser(tBu)-OtBu 954Fmoc-Lys(εFmoc) Leu Arg Thr(tBu)-OtBu 955 Fmoc-Lys(εFmoc) Glu LeuSer(tBu)-OtBu 956 Fmoc-Lys(εFmoc) Glu Leu Thr(tBu)-OtBu 957Boc-Lys(Fmoc) Glu Ile Thr(tBu)-OtBu 958 Boc-Lys(εFmoc) Leu ArgSer(tBu)-OtBu 959 Boc-Lys(εFmoc) Leu Arg Thr(tBu)-OtBu 960Boc-Lys(εFmoc) Glu Leu Ser(tBu)-OtBu 961 Boc-Lys(εFmoc) Glu LeuThr(tBu)-OtBu 962 Boc-Lys(εBoc) Leu Arg Ser(tBu)-OtBu 963 Boc-Lys(εBoc)Arg Phe Thr(tBu)-OtBu 964 Boc-Lys(εBoc) Leu Arg Thr(tBu)-OtBu 965Boc-Lys(εBoc) Glu Ile Thr(tBu) 966 Boc-Lys(εBoc) Glu Val Thr(tBu) 967Boc-Lys(εBoc) Glu Ala Thr(tBu) 968 Boc-Lys(εBoc) Glu Gly Thr(tBu) 969Boc-Lys(εBoc) Glu Leu Ser(tBu)-OtBu 970 Boc-Lys(εBoc) Glu LeuThr(tBu)-OtBu 971

While the peptides of Table 9 are illustrated with particular protectinggroups, it is noted that these groups may be substituted with otherprotecting groups as described herein and/or one or more of the shownprotecting group can be eliminated.

5) Small Peptides Having Either an Acidic or Basic Amino Acid in theCenter Together with a Central Aromatic Amino Acid.

In certain embodiments, the “small” peptides of this invention rangefrom four amino acids to about ten amino acids. The terminal amino acidsare typically hydrophobic either because of a hydrophobic side chain orbecause the terminal amino acids bear one or more hydrophobic protectinggroups end amino acids (X¹ and X⁴) are hydrophobic either because of ahydrophobic side chain or because the side chain or the C and/or Nterminus is blocked with one or more hydrophobic protecting group(s)(e.g., the N-terminus is blocked with Boc-, Fmoc-, Nicotinyl-, etc., andthe C-terminus blocked with (tBu)-OtBu, etc.). Typically, the centralportion of the peptide comprises a basic or acidic amino acid and anaromatic amino acid (e.g., in a 4 mer) or a basic domain or an acidicdomain and an aromatic domain in a longer molecule.

These four-mers can be represented by Formula I in which X¹ and X⁴ arehydrophobic and/or bear hydrophobic protecting group(s) as describedherein and X² is acidic or basic while X³ is aromatic or X² is aromaticwhile X³ is acidic or basic. The peptide can be all L-amino acids orinclude one, or more, or all D-amino acids. Five-mers can be representedby a minor modification of Formula I in which X⁵ is inserted as shown inTable 10 and in which X⁵ is typically an aromatic amino acid.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 10. TABLE 10 Examples of certainpreferred peptides having either an acidic orbasic amino acid in thecenter together with a central aromatic amino acid. SEQ ID X¹ X² X³ X⁵X⁴ NO Fmoc-Lys(εBoc) Arg Trp Tyr(tBu)-OtBu 972 Fmoc-Lys(εBoc) Trp ArgTyr(tBu)-OtBu 973 Fmoc-Lys(εBoc) Arg Tyr Trp-OtBu 974 Fmoc-Lys(εBoc) TyrArg Trp-OtBu 975 Fmoc-Lys(εBoc) Arg Tyr Trp Thr(tBu)-OtBu 976Fmoc-Lys(εBoc) Arg Tyr Thr(tBu)-OtBu 977 Fmoc-Lys(εBoc) Arg TrpThr(tBu)-OtBu 978 Fmoc- Lys(εFmoc) Arg Trp Tyr(tBu)-OtBu 979 Fmoc-Lys(εFmoc) Arg Tyr Trp-OtBu 980 Fmoc- Lys(εFmoc) Arg Tyr TrpThr(tBu)-OtBu 981 Fmoc- Lys(εFmoc) Arg Tyr Thr(tBu)-OtBu 982 Fmoc-Lys(εFmoc) Arg Trp Thr(tBu)-OtBu 983 Boc-Lys(εFmoc) Arg TrpTyr(tBu)-OtBu 984 Boc-Lys(εFmoc) Arg Tyr Trp-OtBu 985 Boc-Lys(εFmoc) ArgTyr Trp Thr(tBu)-OtBu 986 Boc-Lys(εFmoc) Arg Tyr Thr(tBu)-OtBu 987Boc-Lys(εFmoc) Arg Trp Thr(tBu)-OtBu 988 Boc-Glu Lys(εFmoc) ArgTyr(tBu)-OtBu 989 Boc-Lys(εBoc) Arg Trp Tyr(tBu)-OtBu 990 Boc-Lys(εBoc)Arg Tyr Trp-OtBu 991 Boc-Lys(εBoc) Arg Tyr Trp Thr(tBu)-OtBu 992Boc-Lys(εBoc) Arg Tyr Thr(tBu)-OtBu 993 Boc-Lys(εBoc) Arg PheThr(tBu)-OtBu 994 Boc-Lys(εBoc) Arg Trp Thr(tBu)-OtBu 995

While the peptides of Table 10 are illustrated with particularprotecting groups, it is noted that these groups may be substituted withother protecting groups as described herein and/or one or more of theshown protecting group can be eliminated.

6) Small Peptides Having Aromatic Amino Acids or Aromatic Amino AcidsSeparated by Histidine(s) at the Center.

In certain embodiments, the peptides of this invention are characterizedby π electrons that are exposed in the center of the molecule whichallow hydration of the particle and that allow the peptide particles totrap pro-inflammatory oxidized lipids such as fatty acid hydroperoxidesand phospholipids that contain an oxidation product of arachidonic acidat the sn-2 position.

In certain embodiments, these peptides consist of a minimum of 4 aminoacids and a maximum of about 10 amino acids, preferentially (but notnecessarily) with one or more of the amino acids being theD-sterioisomer of the amino acid, with the end amino acids beinghydrophobic either because of a hydrophobic side chain or because theterminal amino acid(s) bear one or more hydrophobic blocking group(s),(e.g., an N-terminus blocked with Boc-, Fmoc-, Nicotinyl-, and the like,and a C-terminus blocked with (tBu)-OtBu groups and the like). Insteadof having an acidic or basic amino acid in the center, these peptidesgenerally have an aromatic amino acid at the center or have aromaticamino acids separated by histidine in the center of the peptide.

Certain preferred peptides of this invention include, but are notlimited to the peptides shown in Table 11. TABLE 11 Examples of peptideshaving aromatic amino acids in the center or aromatic amino acids oraromatic domains separated by one or more histidines. X¹ X² X³ X⁴ X⁵ SEQID NO Boc-Lys(εBoc) Phe Trp Phe Ser(tBu)-OtBu  996 Boc-Lys(εBoc) Phe TrpPhe Thr(tBu)-OtBu  997 Boc-Lys(εBoc) Phe Tyr Phe Ser(tBu)-OtBu  998Boc-Lys(εBoc) Phe Tyr Phe Thr(tBu)-OtBu  999 Boc-Lys(εBoc) Phe His PheSer(tBu)-OtBu 1000 Boc-Lys(εBoc) Phe His Phe Thr(tBu)-OtBu 1001Boc-Lys(εBoc) Val Phe Phe-Tyr Ser(tBu)-OtBu 1002 Nicotinyl-Lys(εBoc) PheTrp Phe Ser(tBu)-OtBu 1003 Nicotinyl-Lys(εBoc) Phe Trp Phe Thr(tBu)-OtBu1004 Nicotinyl-Lys(εBoc) Phe Tyr Phe Ser(tBu)-OtBu 1005Nicotinyl-Lys(εBoc) Phe Tyr Phe Thr(tBu)-OtBu 1006 Nicotinyl-Lys(εBoc)Phe His Phe Ser(tBu)-OtBu 1007 Nicotinyl-Lys(εBoc) Phe His PheThr(tBu)-OtBu 1008 Boc-Leu Phe Trp Phe Thr(tBu)-OtBu 1009 Boc-Leu PheTrp Phe Ser(tBu)-OtBu 1010

While the peptides of Table 11 are illustrated with particularprotecting groups, it is noted that these groups may be substituted withother protecting groups as described herein and/or one or more of theshown protecting group can be eliminated.

7) Summary of Tripeptides and Tetrapeptides.

For the sake of clarity, a number of tripeptides and tetrapeptides ofthis invention are generally summarized below in Table 12. TABLE 12General structure of certain peptides of this invention. X¹ X² X³ X⁴hydrophobic side chain Acidic or — hydrophobic side or hydrophobic Basicchain or protecting group(s) hydrophobic protecting group(s) hydrophobicside chain Basic Acidic hydrophobic side or hydrophobic chain orprotecting group(s) hydrophobic protecting group(s) hydrophobic sidechain Acidic Basic hydrophobic side or hydrophobic chain or protectinggroup(s) hydrophobic protecting group(s) hydrophobic side chain Acidicor Aliphatic hydrophobic side or hydrophobic Basic chain or protectinggroup(s) hydrophobic protecting group(s) hydrophobic side chainAliphatic Acidic or Basic hydrophobic side or hydrophobic chain orprotecting group(s) hydrophobic protecting group(s) hydrophobic sidechain Acidic or Aromatic hydrophobic side or hydrophobic Basic chain orprotecting group(s) hydrophobic protecting group(s) hydrophobic sidechain Aromatic Acidic or Basic hydrophobic side or hydrophobic chain orprotecting group(s) hydrophobic protecting group(s) hydrophobic sidechain Aromatic His Aromatic hydrophobic side or hydrophobic chain orprotecting group(s) hydrophobic protecting group(s)

Where longer peptides are desired, X² and X³ can represent domains(e.g., regions of two or more amino acids of the specified type) ratherthan individual amino acids. Table 12 is intended to be illustrative andnot limiting. Using the teaching provided herein, other suitablepeptides can readily be identified.

8) Paired Amino Acids and Dipeptides.

In certain embodiments, this invention pertains to the discovery thatcertain pairs of amino acids, administered in conjunction with eachother or linked to form a dipeptide have one or more of the propertiesdescribed herein. Thus, without being bound to a particular theory, itis believed that when the pairs of amino acids are administered inconjunction with each other, as described herein, they are capableparticipating in or inducing the formation of micelles in vivo.

Similar to the other small peptides described herein, it is believedthat the pairs of peptides will associate in vivo, and demonstratephysical properties including high solubility in ethyl acetate (e.g.,greater than about 4 mg/mL), solubility in aqueous buffer at pH 7.0.Upon contacting phospholipids such as1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC), in an aqueousenvironment, it is believed the pairs of amino acids induce orparticipate in the formation of particles with a diameter ofapproximately 7.5 nm (±0.1 nm), and/or induce or participate in theformation of stacked bilayers with a bilayer dimension on the order of3.4 to 4.1 nm with spacing between the bilayers in the stack ofapproximately 2 nm, and/or also induce or participate in the formationof vesicular structures of approximately 38 nm).

Moreover, it is further believed that the pairs of amino acids candisplay one or more of the following physiologically relevantproperties:

-   -   1. They convert pro-inflammatory HDL to anti-inflammatory HDL or        make anti-inflammatory HDL more anti-inflammatory;    -   2. They decrease LDL-induced monocyte chemotactic activity        generated by artery wall cells;    -   3. They stimulate the formation and cycling of pre-0 HDL;    -   4. They raise HDL cholesterol; and/or    -   5. They increase HDL paraoxonase activity.

The pairs of amino acids can be administered as separate amino acids(administered sequentially or simultaneously, e.g. in a combinedformulation) or they can be covalently coupled directly or through alinker (e.g. a PEG linker, a carbon linker, a branched linker, astraight chain linker, a heterocyclic linker, a linker formed ofderivatized lipid, etc.). In certain embodiments, the pairs of aminoacids are covalently linked through a peptide bond to form a dipeptide.In various embodiments while the dipeptides will typically comprise twoamino acids each bearing an attached protecting group, this inventionalso contemplates dipeptides wherein only one of the amino acids bearsone or more protecting groups.

The pairs of amino acids typically comprise amino acids where each aminoacid is attached to at least one protecting group (e.g., a hydrophobicprotecting group as described herein). The amino acids can be in the Dor the L form. In certain embodiments, where the amino acids comprisingthe pairs are not attached to each other, each amino acid bears twoprotecting groups (e.g., such as molecules 1 and 2 in Table 13). TABLE13 Illustrative amino acid pairs of this invention. Amino AcidPair/dipeptide 1. Boc-Arg-OtBu* 2. Boc-Glu-OtBu* 3. Boc-Phe-Arg-OtBu**4. Boc-Glu-Leu-OtBu** 5. Boc-Arg-Glu-OtBu****This would typically be administered in conjunction with a second aminoacid.**In certain embodiments, these dipeptides would be administered inconjunction with each other.***In certain embodiments, this peptide would be administered eitheralone or in combination with one of the other peptides describedherein..

Suitable pairs of amino acids can readily be identified by providing thepair of protected amino acids and/or a dipeptide and then screening thepair of amino acids/dipeptide for one or more of the physical and/orphysiological properties described above. In certain embodiments, thisinvention excludes pairs of amino acids and/or dipeptides comprisingaspartic acid and phenylalanine. In certain embodiments, this inventionexcludes pairs of amino acids and/or dipeptides in which one amino acidis (−)-N-[(trans-4-isopropylcyclohexane)carbonyl]-D-phenylalanine(nateglinide).

In certain embodiments, the amino acids comprising the pair areindependently selected from the group consisting of an acidic amino acid(e.g., aspartic acid, glutamic acid, etc.), a basic amino acid (e.g.,lysine, arginine, histidine, etc.), and a non-polar amino acid (e.g.,alanine, valine, leucine, isoleucine, proline, phenylalanine,tryptophan, methionine, etc.). In certain embodiments, where the firstamino acid is acidic or basic, the second amino acid is non-polar andwhere the second amino acid is acidic or basic, the first amino acid isnon-polar. In certain embodiments, where the first amino acid is acidic,the second amino acid is basic, and vice versa. (see, e.g., Table 14).

Similar combinations can be obtained by administering pairs ofdipeptides. Thus, for example in certain embodiments, molecules 3 and 4in Table 13 would be administered in conjunction with each other. TABLE14 Certain generalized amino acid pairs/dipeptides. First Amino acidSecond Amino acid 1. Acidic Basic 2. Basic Acidic 3. Acidic Non-polar 4.Non-polar Acidic 5. Basic Non-polar 6. Non-polar Basic

It is noted that these amino acid pairs/dipeptides are intended to beillustrative and not limiting. Using the teaching provided herein othersuitable amino acid pairs/dipeptides can readily be determined.

E) Apo-J (G* Peptides).

In certain It was a discovery of this invention that peptides thatmimicking the amphipathic helical domains of apo J are capable ofmitigating one or more symptoms of atherosclerosis and/or otherpathologies described herein. Apolipoprotein J possesses a wide nonpolarface termed globular protein-like, or G* amphipathic helical domains.The class G amphipathic helix is found in globular proteins, and thus,the name class G. This class of amphipathic helix is characterized by arandom distribution of positively charged and negatively chargedresidues on the polar face with a narrow nonpolar face. Because of thenarrow nonpolar face this class does not readily associate withphospholipids. The G* of amphipathic helix possesses similar, but notidentical, characteristics to the G amphipathic helix. Similar to theclass G amphipathic helix, the G* class peptides possesses a randomdistribution of positively and negatively charged residues on the polarface. However, in contrast to the class G amphipathic helix which has anarrow nonpolar face, this class has a wide nonpolar face that allowsthis class to readily bind phospholipid and the class is termed G* todifferentiate it from the G class of amphipathic helix.

A number of suitable G* amphipathic peptides are described in copendingapplications U.S. Ser. No. 10/120,508, filed Apr. 5, 2002, U.S. Ser. No.10/520,207, filed Apr. 1, 2003, and PCT Application PCT/US03/09988,filed Apr. 1, 2003. In addition, a variety of suitable peptides of thisinvention that are related to G* amphipathic helical domains of apo Jare illustrated in Table 15. TABLE 15 Certain illustrative peptides foruse in this invention related to G* amphipathic helical domains of apoJ. Amino Acid Sequence SEQ ID NO LLEQLNEQFNWVSRLANLTQGE 1011LLEQLNEQFNWVSRLANL 1012 NELQEMSNQGSKYVNKEIQNAVNGV 1013IQNAVNGVKQIKTLIEKTNEE 1014 RKTLLSNLEEAKKKKEDALNETRESETKLKEL 1015PGVCNETMMALWEECK 1016 PCLKQTCMKFYARVCR 1017 ECKPCLKQTCMKFYARVCR 1018LVGRQLEEFL 1019 MNGDRIDSLLEN 1020 QQTHMLDVMQD 1021 FSRASSIIDELFQD 1022PFLEMIHEAQQAMDI 1023 PTEFIREGDDD 1024 RMKDQCDKCREILSV 1025PSQAKLRRELDESLQVAERLTRKYNELLKSYQ 1026 LLEQLNEQFNWVSRLANLTEGE 1027DQYYLRVTTVA 1028 PSGVTEVVVKLFDS 1029 PKFMETVAEKALQEYRKKHRE 1030

The peptides of this invention, however, are not limited to G* variantsof apo J. Generally speaking G* domains from essentially any otherprotein preferably apo proteins are also suitable. The particularsuitability of such proteins can readily be determined using assays forprotective activity (e.g., protecting LDL from oxidation, and the like),e.g. as illustrated herein in the Examples. Some particularly preferredproteins include G* amphipathic helical domains or variants thereof(e.g., conservative substitutions, and the like) of proteins including,but not limited to apo AI, apo AIV, apo E, apo CII, apo CIII, and thelike.

Certain preferred peptides for related to G* amphipathic helical domainsrelated to apoproteins other than apo J are illustrated in Table 16.TABLE 16 Peptides for use in this invention related to G* amphipathichelical domains related to apoproteins other than apo J. SEQ ID AminoAcid Sequence NO WDRVKDLATVYVDVLKDSGRDYVSQF 1031 (Related to the 8 to 33region of apo AI) VATVMWDYFSQLSNNAKEAVEHLQK 1032 (Related to the 7 to 31region of apo AIV) RWELALGRFWDYLRWVQTLSEQVQEEL 1033 (Related to the 25to 51 region of apo E) LSSQVTQELRALMDETMKELKELKAYKSELEEQLT 1034 (Relatedto the 52 to 83 region of apo E) ARLSKELQAAQARLGADMEDVCGRLV 1035(Related to the 91 to 116 region of apo E) VRLASHLRKLRKRLLRDADDLQKRLA1036 (Related to the 135 to 160 region of apo E) PLVEDMQRQWAGLVEKVQA1037 (267 to 285 of apo E.27) MSTYTGIFTDQVLSVLK 1038 (Related to the 60to 76 region of apo CII) LLSFMQGYMKHATKTAKDALSS 1039 (Related to the 8to 29 region of apo CIII)

Additional illustrative G* peptides are shown in Table 17. TABLE 17Additional illustrative G* peptides. SEQ ID Peptide NOAc-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1040Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1041Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Leu-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1042Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Val-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1043Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Leu-Thr-Glu-Gly-Ser- 1044Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1045Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Ile-Thr-Glu-Gly-Ser- 1046Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Leu-Tyr-His-Val-Thr-Glu-Gly-Ser- 1047Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Val-Tyr-His-Tyr-Thr-Glu-Gly-Ser- 1048Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Phe-Thr-Glu-Gly-Ser- 1049Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Ile-Thr-Glu-Gly-Ser- 1050Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Val-Thr-Glu-Gly-Ser- 1051Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-His-Tyr-Thr-Glu-Gly-Ser- 1052Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Phe-Ile-Trp-His-Leu-Thr-Glu-Gly-Ser- 1053Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Leu-Ile-Trp-His-Leu-Thr-Glu-Gly-Ser- 1054Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Ile-Ile-Trp-His-Leu-Thr-Glu-Gly-Ser- 1055Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Tyr-Ile-Trp-Phe-Leu-Thr-Glu-Gly-Ser- 1056Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-Phe-Leu-Thr-Glu-Gly-Ser- 1057Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-Leu-Leu-Thr-Glu-Gly-Ser- 1058Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1059Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Tyr-Thr-Glu-Gly-Ser- 1060Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Ile-Thr-Glu-Gly-Ser- 1061Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Ser-Glu-Gly-Ser- 1062Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Asp-Gly-Ser- 1063Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Thr- 1064Ser-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1065Thr-Glu-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1066Thr-Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1067Thr-Asp-Tyr-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1068Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1069Thr-Asp-Val-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1070Thr-Asp-Leu-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1071Thr-Asp-Leu-Arg-Ser-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1072Thr-Asp-Leu-Arg-Thr-Asp-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1073Thr-Asp-Ile-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1074Thr-Asp-Ile-Arg-Ser-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1075Thr-Asp-Ile-Lys-Ser-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1076Thr-Asp-Ile-Lys-Ser-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1077Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Tyr-Ile-Trp-His-Leu-Thr-Glu-Gly-Ser- 1078Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1079Thr-Asp-Ile-Arg-Thr-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Phe-His-Leu-Thr-Glu-Gly-Ser- 1080Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1081Thr-Asp-Leu-Lys-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Asp-Gly-Ser- 1082Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Asp-Gly-Ser- 1083Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-Phe-Leu-Thr-Glu-Gly-Ser- 1084Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-Phe-Leu-Thr-Glu-Gly-Ser- 1085Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1086Thr-Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1087Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Phe-His-Leu-Thr-Glu-Gly-Ser- 1088Thr-Asp-Ile-Arg-Thr-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1089Thr-Asp-Ile-Arg-Thr-Asp-Gly-NH₂Ac-Arg-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1090Thr-Asp-Leu-Arg-Thr-Asp-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1091Thr-Asp-Ile-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1092Thr-Asp-Ile-Lys-Thr-Asp-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1093Thr-Asp-Phe-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1094Thr-Asp-Tyr-Lys-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Ile-Tyr-His-Leu-Thr-Glu-Gly-Ser- 1095Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1096Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1097Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1098Thr-Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Lys-Trp-Phe-Tyr-His-Phe-Thr-Asp-Gly-Ser- 1099Thr-Asp-Ile-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1100Thr-Asp-Leu-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1101Thr-Asp-Phe-Arg-Thr-Glu-Gly-NH₂Ac-Arg-Trp-Phe-Tyr-His-Phe-Thr-Glu-Gly-Ser- 1102Thr-Asp-Phe-Arg-Thr-Asp-Gly-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Leu- 1103Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Leu- 1104Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Asp-Glu-Phe-Lys-Ser-Leu- 1105Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Asp-Phe-Lys-Ser-Leu- 1106Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Leu- 1107Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Cys-Val-Asp-Asp-Phe-Lys-Ser-Leu- 1108Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Leu- 1109Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Asp-Asp-Phe-Lys-Ser-Leu- 1110Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1111Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Ile- 1112Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Val- 1113Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Tyr- 1114Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1115Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Ile- 1116Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Val- 1117Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Tyr- 1118Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1119Thr-Thr-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Ile- 1120Ser-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Val- 1121Ser-Thr-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Tyr- 1122Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1123Thr-Thr-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1124Ser-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1125Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1126Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1127Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1128Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1129Thr-Ser-Cys-Phe-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1130Thr-Ser-Cys-Leu-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1131Thr-Ser-Cys-Ile-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Leu-Lys-Ser-Phe- 1132Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1133Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1134Thr-Ser-Cys-Phe-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1135Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1136Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1137Thr-Ser-Cys-Phe-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1138Ser-Ser-Cys-Phe-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1139Gln-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1140Gln-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Gln-Phe- 1141Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Gln-Leu- 1142Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1143Gln-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Gln-Phe- 1144Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1145Thr-Ser-Cys-Phe-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1146Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1147Thr-Ser-Cys-Phe-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Arg-Cys-Val-Glu-Glu-Phe-Lys-Ser-Leu- 1148Thr-Ser-Cys-Leu-Glu-Ser-Lys-Ala-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Leu- 1149Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1150Thr-Ser-Cys-Phe-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1151Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1152Thr-Ser-Cys-Leu-Glu-Ser-Lys-Phe-Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Leu-Lys-Ser-Phe- 1153Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Arg-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1154Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1155Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1156Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1157Thr-Ser-Ala-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Ala-Val-Glu-Glu-Phe-Lys-Ser-Phe- 1158Thr-Ser-Ala-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Arg-Ala-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1159Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Asp-Arg-Ala-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1160Thr-Ser-Ala-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Phe- 1161Thr-Ser-Cys-Phe-Glu-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Cys-Tyr-Glu-Glu-Phe-Lys-Ser-Phe- 1162Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Asp-Lys-Cys-Trp-Glu-Glu-Phe-Lys-Ser-Phe- 1163Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Tyr- 1164Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Trp- 1165Thr-Ser-Cys-Leu-Asp-Ser-Lys-Phe-Phe-NH₂Ac-Glu-Lys-Cys-Val-Glu-Glu-Phe-Lys-Ser-Trp- 1166Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂Ac-Asp-Lys-Cys-Phe-Glu-Glu-Phe-Lys-Ser-Trp- 1167Thr-Ser-Cys-Leu-Asp-Ser-Lys-Ala-Phe-NH₂

Other suitable peptides include, but are not limited to the peptides ofTable 18. TABLE 18 Illustrative peptides having an improved hydrophobicphase. SEQ ID Name Sequence NO V2W3A5F1017- Ac-Asp-Val-Trp-Lys-Ala-Ala-1168 D-4F Tyr-Asp-Lys-Phe-Ala-Glu-Lys- Phe-Lys-Glu-Phe-Phe-NH₂V2W3F10-D-4F Ac-Asp-Val-Trp-Lys-Ala-Phe- 1169Tyr-Asp-Lys-Phe-Ala-Glu-Lys- Phe-Lys-Glu-Ala-Phe-NH₂ W3-D-4FAc-Asp-Phe-Trp-Lys-Ala-Phe- 1170 Tyr-Asp-Lys-Val-Ala-Glu-Lys-Phe-Lys-Glu-Ala-Phe-NH₂

The peptides described here (V2W3A5F10,17-D-4F; V2W3F10-D-4F; W3-D-4F)may be more potent than the original D-4F.

Still other suitable peptides include, but are not limited to:P¹-Dimethyltyrosine-D-Arg-Phe-Lys-P² (SEQ ID NO:1171) andP¹-Dimethyltyrosine-Arg-Glu-Leu-P² where P1 and P2 are protecting groupsas described herein. In certain embodiments, these peptides include, butare not limited to BocDimethyltyrosine-D-Arg-Phe-Lys(OtBu) andBocDimethyltyrosine-Arg-Glu-Leu(OtBu).

In certain embodiments, the peptides of this invention include peptidescomprising or consisting of the amino acid sequence LAEYHAK (SEQ ID NO:1172) comprising at least one D amino acid and/or at least one or twoterminal protecting groups. In certain embodiments, this inventionincludes a peptide that ameliorates one or more symptoms of aninflammatory condition, wherein the peptide: ranges in length from about3 to about 10 amino acids; comprises an amino acid sequence where thesequence comprises acidic or basic amino acids alternating with aromaticor hydrophobic amino acids; comprises hydrophobic terminal amino acidsor terminal amino acids bearing a hydrophobic protecting group; is notthe sequence LAEYHAK (SEQ ID NO: 1173) comprising all L amino acids;where the peptide converts pro-inflammatory HDL to anti-inflammatory HDLand/or makes anti-inflammatory HDL more anti-inflammatory.

It is also noted that the peptides listed in the Tables herein are notfully inclusive. Using the teaching provided herein, other suitablepeptides can routinely be produced (e.g. by conservative orsemi-conservative substitutions (e.g. D replaced by E), extensions,deletions, and the like). Thus, for example, one embodiment utilizestruncations of any one or more of peptides identified by SEQ IDNos:1011-1039.

Longer peptides are also suitable. Such longer peptides may entirelyform a class G or G* amphipathic helix, or the G amphipathic helix(helices) can form one or more domains of the peptide. In addition, thisinvention contemplates multimeric versions of the peptides. Thus, forexample, the peptides illustrated in the tables herein can be coupledtogether (directly or through a linker (e.g. a carbon linker, or one ormore amino acids) with one or more intervening amino acids). Suitablelinkers include, but are not limited to Proline (-Pro-), Gly₄Ser₃ (SEQID NO: 1174), and the like. Thus, one illustrative multimeric peptideaccording to this invention is (D-J336)-P-(D-J336) (i.e.Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-P-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-NH₂,SEQ ID NO: 1175).

This invention also contemplates the use of “hybrid” peptides comprisinga one or more G or G* amphipathic helical domains and one or more classA amphipathic helices. Suitable class A amphipathic helical peptides aredescribed in PCT publication WO 02/15923. Thus, by way of illustration,one such “hybrid” peptide is (D-J336)-Pro-(4F) (i.e.Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂,SEQ ID NO: 1176), and the like.

Using the teaching provided herein, one of skill can routinely modifythe illustrated amphipathic helical peptides to produce other suitableapo J variants and/or amphipathic G and/or A helical peptides of thisinvention. For example, routine conservative or semi-conservativesubstitutions (e.g., E for D) can be made of the existing amino acids.The effect of various substitutions on lipid affinity of the resultingpeptide can be predicted using the computational method described byPalgunachari et al. (1996) Arteriosclerosis, Thrombosis, & VascularBiology 16: 328-338. The peptides can be lengthened or shortened as longas the class helix structure(s) are preserved. In addition,substitutions can be made to render the resulting peptide more similarto peptide(s) endogenously produced by the subject species.

While, in preferred embodiments, the peptides of this invention utilizenaturally-occurring amino acids or D forms of naturally occurring aminoacids, substitutions with non-naturally occurring amino acids (e.g.,methionine sulfoxide, methionine methylsulfonium, norleucine,episilon-aminocaproic acid, 4-aminobutanoic acid,tetrahydroisoquinoline-3-carboxylic acid, 8-aminocaprylic acid,4-aminobutyric acid, Lys(N(epsilon)-trifluoroacetyl), α-aminoisobutyricacid, and the like) are also contemplated.

New peptides can be designed and/or evaluated using computationalmethods. Computer programs to identify and classify amphipathic helicaldomains are well known to those of skill in the art and many have beendescribed by Jones et al. (1992) J. Lipid Res. 33: 287-296). Suchprograms include, but are not limited to the helical wheel program(WHEEL or WHEEL/SNORKEL), helical net program (HELNET, HELNET/SNORKEL,HELNET/Angle), program for addition of helical wheels (COMBO orCOMBO/SNORKEL), program for addition of helical nets (COMNET,COMNET/SNORKEL, COMBO/SELECT, COMBO/NET), consensus wheel program(CONSENSUS, CONSENSUS/SNORKEL), and the like.

F) Blocking Groups and D Residues.

While the various peptides and/or amino acid pairs described herein maybe shown with no protecting groups, in certain embodiments (e.g.particularly for oral administration), they can bear one, two, three,four, or more protecting groups. The protecting groups can be coupled tothe C- and/or N-terminus of the peptide(s) and/or to one or moreinternal residues comprising the peptide(s) (e.g., one or more R-groupson the constituent amino acids can be blocked). Thus, for example, incertain embodiments, any of the peptides described herein can bear, e.g.an acetyl group protecting the amino terminus and/or an amide groupprotecting the carboxyl terminus. One example of such a “dual protectedpeptide is Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-NH₂ (SEQ IDNO:1011 with blocking groups), either or both of these protecting groupscan be eliminated and/or substituted with another protecting group asdescribed herein.

Without being bound by a particular theory, it was a discovery of thisinvention that blockage, particularly of the amino and/or carboxyltermini of the subject peptides of this invention greatly improves oraldelivery and significantly increases serum half-life.

A wide number of protecting groups are suitable for this purpose. Suchgroups include, but are not limited to acetyl, amide, and alkyl groupswith acetyl and alkyl groups being particularly preferred for N-terminalprotection and amide groups being preferred for carboxyl terminalprotection. In certain particularly preferred embodiments, theprotecting groups include, but are not limited to alkyl chains as infatty acids, propeonyl, formyl, and others. Particularly preferredcarboxyl protecting groups include amides, esters, and ether-formingprotecting groups. In one preferred embodiment, an acetyl group is usedto protect the amino terminus and an amide group is used to protect thecarboxyl terminus. These blocking groups enhance the helix-formingtendencies of the peptides. Certain particularly preferred blockinggroups include alkyl groups of various lengths, e.g. groups having theformula: CH₃—(CH₂)_(n)—CO— where n ranges from about 1 to about 20,preferably from about 1 to about 16 or 18, more preferably from about 3to about 13, and most preferably from about 3 to about 10.

In certain particularly preferred embodiments, the protecting groupsinclude, but are not limited to alkyl chains as in fatty acids,propeonyl, formyl, and others. Particularly preferred carboxylprotecting groups include amides, esters, and ether-forming protectinggroups. In one preferred embodiment, an acetyl group is used to protectthe amino terminus and an amide group is used to protect the carboxylterminus. These blocking groups enhance the helix-forming tendencies ofthe peptides. Certain particularly preferred blocking groups includealkyl groups of various lengths, e.g. groups having the formula:CH₃—(CH₂)_(n)—CO— where n ranges from about 3 to about 20, preferablyfrom about 3 to about 16, more preferably from about 3 to about 13, andmost preferably from about 3 to about 10.

Other protecting groups include, but are not limited to Fmoc,t-butoxycarbonyl (t-BOC), 9-fluoreneacetyl group, 1-fluorenecarboxylicgroup, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl(MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl),Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z),2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom), cyclohexyloxy(cHxO), t-butoxymethyl (Bum), t-butoxy (tBuO), t-Butyl (tBu), Acetyl(Ac), and Trifluoroacetyl (TFA).

Protecting/blocking groups are well known to those of skill as aremethods of coupling such groups to the appropriate residue(s) comprisingthe peptides of this invention (see, e.g., Greene et al., (1991)Protective Groups in Organic Synthesis, 2nd ed., John Wiley & Sons, Inc.Somerset, N.J.). In one preferred embodiment, for example, acetylationis accomplished during the synthesis when the peptide is on the resinusing acetic anhydride. Amide protection can be achieved by theselection of a proper resin for the synthesis. During the synthesis ofthe peptides described herein in the examples, rink amide resin wasused. After the completion of the synthesis, the semipermanentprotecting groups on acidic bifunctional amino acids such as Asp and Gluand basic amino acid Lys, hydroxyl of Tyr are all simultaneouslyremoved. The peptides released from such a resin using acidic treatmentcomes out with the n-terminal protected as acetyl and the carboxylprotected as NH₂ and with the simultaneous removal of all of the otherprotecting groups.

In certain particularly preferred embodiments, the peptides comprise oneor more D-form (dextro rather than levo) amino acids as describedherein. In certain embodiments at least two enantiomeric amino acids,more preferably at least 4 enantiomeric amino acids and most preferablyat least 8 or 10 enantiomeric amino acids are “D” form amino acids. Incertain embodiments every other, ore even every amino acid (e.g. everyenantiomeric amino acid) of the peptides described herein is a D-formamino acid.

In certain embodiments at least 50% of the enantiomeric amino acids are“D” form, more preferably at least 80% of the enantiomeric amino acidsare “D” form, and most preferably at least 90% or even all of theenantiomeric amino acids are “D” form amino acids.

G) Peptide Mimetics.

In addition to the peptides described herein, peptidomimetics are alsocontemplated. Peptide analogs are commonly used in the pharmaceuticalindustry as non-peptide drugs with properties analogous to those of thetemplate peptide. These types of non-peptide compound are termed“peptide mimetics” or “peptidomimetics” (Fauchere (1986) Adv. Drug Res.15: 29; Veber and Freidinger (1985) TINS p. 392; and Evans et al. (1987)J. Med. Chem. 30: 1229) and are usually developed with the aid ofcomputerized molecular modeling. Peptide mimetics that are structurallysimilar to therapeutically useful peptides may be used to produce anequivalent therapeutic or prophylactic effect.

Generally, peptidomimetics are structurally similar to a paradigmpolypeptide (e.g. SEQ ID NO:5 shown in Table 1), but have one or morepeptide linkages optionally replaced by a linkage selected from thegroup consisting of: —CH₂NH—, —CH₂S—, —CH₂—CH₂—, —CH═CH— (cis andtrans), —COCH₂—, —CH(OH)CH₂—, —CH₂SO—, etc. by methods known in the artand further described in the following references: Spatola (1983) p. 267in Chemistry and Biochemistry of Amino Acids, Peptides, and Proteins, B.Weinstein, eds., Marcel Dekker, New York,; Spatola (1983) Vega Data 1(3)Peptide Backbone Modifications. (general review); Morley (1980) TrendsPharm Sci pp. 463-468 (general review); Hudson et al. (1979) Int J PeptProt Res 14:177-185 (—CH₂NH—, CH₂CH₂—); Spatola et al. (1986) Life Sci38:1243-1249 (—CH₂—S); Hann, (1982) J Chem Soc Perkin Trans 1307-314(—CH—CH—, cis and trans); Almquist et al. (1980) J Med. Chem.23:1392-1398 (—COCH₂—); Jennings-White et al. (1982) Tetrahedron Lett.23:2533 (—COCH₂—); Szelke et al., European Appln. EP 45665 (1982) CA:97:39405 (1982) (—CH(OH)CH2-); Holladay et al. (1983) Tetrahedron Lett24:4401-4404 (—C(OH)CH₂—); and Hruby (1982) Life Sci., 31:189-199(—CH₂—S—)).

One particularly preferred non-peptide linkage is —CH₂NH—. Such peptidemimetics may have significant advantages over polypeptide embodiments,including, for example: more economical production, greater chemicalstability, enhanced pharmacological properties (half-life, absorption,potency, efficacy, etc.), reduced antigenicity, and others.

In addition, circularly permutations of the peptides described herein orconstrained peptides (including cyclized peptides) comprising aconsensus sequence or a substantially identical consensus sequencevariation may be generated by methods known in the art (Rizo andGierasch (1992) Ann. Rev. Biochem. 61: 387); for example, by addinginternal cysteine residues capable of forming intramolecular disulfidebridges which cyclize the peptide.

H) Small Organic Molecules.

In certain embodiments, the active agents of this invention includesmall organic molecules, e.g. as described in copending application U.S.Ser. No. 60/600,925, filed Aug. 11, 2004. In various embodiments thesmall organic molecules are similar to, and in certain cases, mimeticsof the tetra- and penta-peptides described in copending application U.S.Ser. No. 10/649,378, filed on Aug. 26, 2003 and U.S. Ser. No.60/494,449, filed on August 11.

The small organic molecules of this invention typically have molecularweights less than about 900 Daltons. Typically the molecules are highlysoluble in ethyl acetate (e.g., at concentrations equal to or greaterthan 4 mg/mL), and also are soluble in aqueous buffer at pH 7.0.

Contacting phospholipids such as1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), with the smallorganic molecules of this invention in an aqueous environment typicallyresults in the formation of particles with a diameter of approximately7.5 nm (±0.1 nm). In addition, stacked bilayers are often formed with abilayer dimension on the order of 3.4 to 4.1 nm with spacing between thebilayers in the stack of approximately 2 nm. Vesicular structures ofapproximately 38 nm are also often formed. Moreover, when the moleculesof this invention are administered to a mammal they render HDL moreanti-inflammatory and mitigate one or more symptoms of atherosclerosisand/or other conditions characterized by an inflammatory response.

Thus, in certain embodiments, the small organic molecule is one thatameliorates one or more symptoms of a pathology characterized by aninflammatory response in a mammal (e.g. atherosclerosis), where thesmall molecule is soluble in ethyl acetate at a concentration greaterthan 4 mg/mL, is soluble in aqueous buffer at pH 7.0, and, whencontacted with a phospholipid in an aqueous environment, forms particleswith a diameter of approximately 7.5 nm and forms stacked bilayers witha bilayer dimension on the order of 3.4 to 4.1 nm with spacing betweenthe bilayers in the stack of approximately 2 nm, and has a molecularweightless than 900 daltons.

In certain embodiment, the molecule has the formula:

where P¹, P², P³, and P⁴ are independently selected hydrophobicprotecting groups; R¹ and R⁴ are independently selected amino acid Rgroups; n, i, x, y, and z are independently zero or 1 such that when nand x are both zero, R¹ is a hydrophobic group and when y and i are bothzero, R⁴ is a hydrophobic group; R² and R³ are acidic or basic groups atpH 7.0 such that when R² is acidic, R³ is basic and when R² is basic, R³is acidic; and R⁵, when present is selected from the group consisting ofan aromatic group, an aliphatic group, a positively charged group, or anegatively charged group. In certain embodiments, R² or R³ is—(CH₂)j-COOH where j=1, 2, 3, or 4 and/or —(CH₂)j-NH₂ where j=1, 2, 3,4, or 5, or —(CH₂)j-NH—C(═NH)—NH₂ where n=1, 2, 3 or 4. In certainembodiments, R², R³, and R⁵, when present, are amino acid R groups.Thus, for example, In various embodiments R² and R³ are independently anaspartic acid R group, a glutamic acid R group, a lysine R group, ahistidine R group, or an arginine R group (e.g., as illustrated in Table1).

In certain embodiments, R¹ is selected from the group consisting of aLys R group, a Trp R group, a Phe R group, a Leu R group, an Orn Rgroup, pr a norLeu R group. In certain embodiments, R⁴ is selected fromthe group consisting of a Ser R group, a Thr R group, an Ile R group, aLeu R group, a norLeu R group, a Phe R group, or a Tyr R group.

In various embodiments x is 1, and R⁵ is an aromatic group (e.g., a TrpR group).

In various embodiments at least one of n, x, y, and i is 1 and P¹, P²,P³, and P⁴ when present, are independently selected from the groupconsisting of polyethylene glycol (PEG), an acetyl, amide, a 3 to 20carbon alkyl group, fmoc, 9-fluoreneacetyl group, 1-fluorenecarboxylicgroup, 9-fluorenecarboxylic, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), -4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl(MeBzl), 4-methoxybenzyl (MeOBzl), benzyloxy (BzlO), benzyl (Bzl),benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z),2-bromobenzyloxycarbonyl (2-Br-Z), benzyloxymethyl (Bom),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), a propyl group, a butyl group, a pentylgroup, a hexyl group, and trifluoroacetyl (TFA). In certain embodiments,P¹ when present and/or P² when present are independently selected fromthe group consisting of Boc-, Fmoc-, and Nicotinyl- and/or P³ whenpresent and/or P⁴ when present are independently selected from the groupconsisting of tBu, and OtBu.

While a number of protecting groups (P¹, P², P³, P⁴) are illustratedabove, this list is intended to be illustrative and not limiting. Inview of the teachings provided herein, a number of otherprotectingiblocking groups will also be known to one of skill in theart. Such blocking groups can be selected to minimize digestion (e.g.,for oral pharmaceutical delivery), and/or to increaseuptake/bioavailability (e.g., through mucosal surfaces in nasaldelivery, inhalation therapy, rectal administration), and/or to increaseserum/plasma half-life. In certain embodiments, the protecting groupscan be provided as an excipient or as a component of an excipient.

In certain embodiments, z is zero and the molecule has the formula:

where P¹, P², P³, P⁴, R¹, R², R³, R⁴, n, x, y, and i are as describedabove.

In certain embodiments, z is zero and the molecule has the formula:

where R¹, R², R³, and R⁴ are as described above.

In one embodiment, the molecule has the formula:

In certain embodiments, this invention contemplates small moleculeshaving one or more of the physical and/or functional propertiesdescribed herein and having the formula:

where P¹, P², P³, and P⁴ are independently selected hydrophobicprotecting groups as described above, n, x, and y are independently zeroor 1; j, k, and 1 are independently zero, 1, 2, 3, 4, or 5; and R² andR³ are acidic or basic groups at pH 7.0 such that when R² is acidic, R³is basic and when R² is basic, R³ is acidic. In certain preferredembodiments, the small molecule is soluble in water; and the smallmolecule has a molecular weight less than about 900 Daltons. In certainembodiments, n, x, y, j, and 1 are 1; and k is 4.

In certain embodiments, P¹ and/or P² are aromatic protecting groups. Incertain embodiments, R² and R³ are amino acid R groups, e.g., asdescribed above. In various embodiments least one of n, x, and y, is 1and P¹, P², P³ and P⁴ when present, are independently protecting groups,e.g. as described above. In certain embodiments the protecting groups,when present, are independently selected from the group consisting ofpolyethylene glycol (PEG), an acetyl, amide, 3 to 20 carbon alkylgroups, Fmoc, 9-fluoreneacetyl group, 1-fluorenecarboxylic group,9-fluorenecarboxylic, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), -4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-penta

III. Functional Assays of Active Agents.

Certain active agents for use in the methods of this invention aredescribed herein by various formulas (e.g., Formula I, above) and/or byparticular sequences. In certain embodiments, preferred active agents ofthis invention are characterized by one or more of the followingfunctional properties:

-   -   1. They convert pro-inflammatory HDL to anti-inflammatory HDL or        make anti-inflammatory HDL more anti-inflammatory;    -   2. They decrease LDL-induced monocyte chemotactic activity        generated by artery wall cells;    -   3. They stimulate the formation and cycling of pre-0 HDL;    -   4. They raise HDL cholesterol; and/or    -   5. They increase HDL paraoxonase activity.

The specific agents disclosed herein, and/or agents corresponding to thevarious formulas described herein can readily be tested for one or moreof these activities as desired.

Methods of screening for each of these functional properties are wellknown to those of skill in the art. In particular, it is noted thatassays for monocyte chemotactic activity, HDL cholesterol, and HDL HDLparaoxonase activity are illustrated in PCT/US01/26497 (WO 2002/15923).

IV. Peptide Preparation.

The peptides used in this invention can be chemically synthesized usingstandard chemical peptide synthesis techniques or, particularly wherethe peptide does not comprise “D” amino acid residues, can berecombinantly expressed. In certain embodiments, even peptidescomprising “D” amino acid residues are recombinantly expressed. Wherethe polypeptides are recombinantly expressed, a host organism (e.g.bacteria, plant, fungal cells, etc.) in cultured in an environment whereone or more of the amino acids is provided to the organism exclusivelyin a D form. Recombinantly expressed peptides in such a system thenincorporate those D amino acids.

In preferred embodiments the peptides are chemically synthesized by anyof a number of fluid or solid phase peptide synthesis techniques knownto those of skill in the art. Solid phase synthesis in which theC-terminal amino acid of the sequence is attached to an insolublesupport followed by sequential addition of the remaining amino acids inthe sequence is a preferred method for the chemical synthesis of thepolypeptides of this invention. Techniques for solid phase synthesis arewell known to those of skill in the art and are described, for example,by Barany and Merrifield (1963) Solid-Phase Peptide Synthesis; pp. 3-284in The Peptides: Analysis, Synthesis, Biology. Vol. 2: Special Methodsin Peptide Synthesis, Part A; Merrifield et al. (1963) J. Am. Chem.Soc., 85: 2149-2156, and Stewart et al. (1984) Solid Phase PeptideSynthesis, 2nd ed. Pierce Chem. Co., Rockford, Ill.

In certain embodiments, the peptides are synthesized by the solid phasepeptide synthesis procedure using a benzhyderylamine resin (BeckmanBioproducts, 0.59 mmol of NH₂/g of resin) as the solid support. The COOHterminal amino acid (e.g., t-butylcarbonyl-Phe) is attached to the solidsupport through a 4-(oxymethyl)phenacetyl group. This is a more stablelinkage than the conventional benzyl ester linkage, yet the finishedpeptide can still be cleaved by hydrogenation. Transfer hydrogenationusing formic acid as the hydrogen donor is used for this purpose.Detailed protocols used for peptide synthesis and analysis ofsynthesized peptides are described in a miniprint supplementaccompanying Anantharamaiah et al. (1985) J. Biol. Chem., 260(16):10248-10255.

It is noted that in the chemical synthesis of peptides, particularlypeptides comprising D amino acids, the synthesis usually produces anumber of truncated peptides in addition to the desired full-lengthproduct. The purification process (e.g. HPLC) typically results in theloss of a significant amount of the full-length product.

It was a discovery of this invention that, in the synthesis of a Dpeptide (e.g. D-4), in order to prevent loss in purifying the longestform one can dialyze and use the mixture and thereby eliminate the lastHPLC purification. Such a mixture loses about 50% of the potency of thehighly purified product (e.g. per wt of protein product), but themixture contains about 6 times more peptide and thus greater totalactivity.

V. Pharmaceutical Formulations and Devices.

A) Pharmaceutical Formulations.

In order to carry out the methods of the invention, one or more activeagents of this invention are administered, e.g. to an individualdiagnosed as having one or more symptoms of atherosclerosis, or as beingat risk for atherosclerosis and or the various other pathologiesdescribed herein. The active agent(s) can be administered in the“native” form or, if desired, in the form of salts, esters, amides,prodrugs, derivatives, and the like, provided the salt, ester, amide,prodrug or derivative is suitable pharmacologically, i.e., effective inthe present method. Salts, esters, amides, prodrugs and otherderivatives of the active agents can be prepared using standardprocedures known to those skilled in the art of synthetic organicchemistry and described, for example, by March (1992) Advanced OrganicChemistry; Reactions, Mechanisms and Structure, 4th Ed. N.Y.Wiley-Interscience.

For example, acid addition salts are prepared from the free base usingconventional methodology, that typically involves reaction with asuitable acid. Generally, the base form of the drug is dissolved in apolar organic solvent such as methanol or ethanol and the acid is addedthereto. The resulting salt either precipitates or can be brought out ofsolution by addition of a less polar solvent. Suitable acids forpreparing acid addition salts include both organic acids, e.g., aceticacid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malicacid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaricacid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like, as well as inorganic acids, e.g.,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. An acid addition salt may be reconvertedto the free base by treatment with a suitable base. Particularlypreferred acid addition salts of the active agents herein are halidesalts, such as may be prepared using hydrochloric or hydrobromic acids.Conversely, preparation of basic salts of the active agents of thisinvention are prepared in a similar manner using a pharmaceuticallyacceptable base such as sodium hydroxide, potassium hydroxide, ammoniumhydroxide, calcium hydroxide, trimethylamine, or the like. Particularlypreferred basic salts include alkali metal salts, e.g., the sodium salt,and copper salts.

Preparation of esters typically involves functionalization of hydroxyland/or carboxyl groups which may be present within the molecularstructure of the drug. The esters are typically acyl-substitutedderivatives of free alcohol groups, i.e., moieties that are derived fromcarboxylic acids of the formula RCOOH where R is alky, and preferably islower alkyl. Esters can be reconverted to the free acids, if desired, byusing conventional hydrogenolysis or hydrolysis procedures.

Amides and prodrugs can also be prepared using techniques known to thoseskilled in the art or described in the pertinent literature. Forexample, amides may be prepared from esters, using suitable aminereactants, or they may be prepared from an anhydride or an acid chlorideby reaction with ammonia or a lower alkyl amine. Prodrugs are typicallyprepared by covalent attachment of a moiety that results in a compoundthat is therapeutically inactive until modified by an individual'smetabolic system.

The active agents identified herein are useful for parenteral, topical,oral, nasal (or otherwise inhaled), rectal, or local administration,such as by aerosol or transdermally, for prophylactic and/or therapeutictreatment of one or more of the pathologies/indications described herein(e.g., atherosclerosis and/or symptoms thereof). The pharmaceuticalcompositions can be administered in a variety of unit dosage formsdepending upon the method of administration. Suitable unit dosage forms,include, but are not limited to powders, tablets, pills, capsules,lozenges, suppositories, patches, nasal sprays, injectibles, implantablesustained-release formulations, lipid complexes, etc.

The active agents of this invention are typically combined with apharmaceutically acceptable carrier (excipient) to form apharmacological composition. Pharmaceutically acceptable carriers cancontain one or more physiologically acceptable compound(s) that act, forexample, to stabilize the composition or to increase or decrease theabsorption of the active agent(s). Physiologically acceptable compoundscan include, for example, carbohydrates, such as glucose, sucrose, ordextrans, antioxidants, such as ascorbic acid or glutathione, chelatingagents, low molecular weight proteins, protection and uptake enhancerssuch as lipids, compositions that reduce the clearance or hydrolysis ofthe active agents, or excipients or other stabilizers and/or buffers.

Other physiologically acceptable compounds include wetting agents,emulsifying agents, dispersing agents or preservatives that areparticularly useful for preventing the growth or action ofmicroorganisms. Various preservatives are well known and include, forexample, phenol and ascorbic acid. One skilled in the art wouldappreciate that the choice of pharmaceutically acceptable carrier(s),including a physiologically acceptable compound depends, for example, onthe route of administration of the active agent(s) and on the particularphysio-chemical characteristics of the active agent(s).

The excipients are preferably sterile and generally free of undesirablematter. These compositions may be sterilized by conventional, well-knownsterilization techniques.

In therapeutic applications, the compositions of this invention areadministered to a patient suffering from one or more symptoms of the oneor more pathologies described herein, or at risk for one or more of thepathologies described herein in an amount sufficient to prevent and/orcure and/or or at least partially prevent or arrest the disease and/orits complications. An amount adequate to accomplish this is defined as a“therapeutically effective dose.” Amounts effective for this use willdepend upon the severity of the disease and the general state of thepatient's health. Single or multiple administrations of the compositionsmay be administered depending on the dosage and frequency as requiredand tolerated by the patient. In any event, the composition shouldprovide a sufficient quantity of the active agents of the formulationsof this invention to effectively treat (ameliorate one or more symptoms)the patient.

The concentration of active agent(s) can vary widely, and will beselected primarily based on fluid volumes, viscosities, body weight andthe like in accordance with the particular mode of administrationselected and the patient's needs. Concentrations, however, willtypically be selected to provide dosages ranging from about 0.1 or 1mg/kg/day to about 50 mg/kg/day and sometimes higher. Typical dosagesrange from about 3 mg/kg/day to about 3.5 mg/kg/day, preferably fromabout 3.5 mg/kg/day to about 7.2 mg/kg/day, more preferably from about7.2 mg/kg/day to about 11.0 mg/kg/day, and most preferably from about11.0 mg/kg/day to about 15.0 mg/kg/day. In certain preferredembodiments, dosages range from about 10 mg/kg/day to about 50mg/kg/day. In certain embodiments, dosages range from about 20 mg toabout 50 mg given orally twice daily. It will be appreciated that suchdosages may be varied to optimize a therapeutic regimen in a particularsubject or group of subjects.

In certain preferred embodiments, the active agents of this inventionare administered orally (e.g. via a tablet) or as an injectable inaccordance with standard methods well known to those of skill in theart. In other preferred embodiments, the peptides, may also be deliveredthrough the skin using conventional transdermal drug delivery systems,i.e., transdermal “patches” wherein the active agent(s) are typicallycontained within a laminated structure that serves as a drug deliverydevice to be affixed to the skin. In such a structure, the drugcomposition is typically contained in a layer, or “reservoir,”underlying an upper backing layer. It will be appreciated that the term“reservoir” in this context refers to a quantity of “activeingredient(s)” that is ultimately available for delivery to the surfaceof the skin. Thus, for example, the “reservoir” may include the activeingredient(s) in an adhesive on a backing layer of the patch, or in anyof a variety of different matrix formulations known to those of skill inthe art. The patch may contain a single reservoir, or it may containmultiple reservoirs.

In one embodiment, the reservoir comprises a polymeric matrix of apharmaceutically acceptable contact adhesive material that serves toaffix the system to the skin during drug delivery. Examples of suitableskin contact adhesive materials include, but are not limited to,polyethylenes, polysiloxanes, polyisobutylenes, polyacrylates,polyurethanes, and the like. Alternatively, the drug-containingreservoir and skin contact adhesive are present as separate and distinctlayers, with the adhesive underlying the reservoir which, in this case,may be either a polymeric matrix as described above, or it may be aliquid or hydrogel reservoir, or may take some other form. The backinglayer in these laminates, which serves as the upper surface of thedevice, preferably functions as a primary structural element of the“patch” and provides the device with much of its flexibility. Thematerial selected for the backing layer is preferably substantiallyimpermeable to the active agent(s) and any other materials that arepresent.

Other preferred formulations for topical drug delivery include, but arenot limited to, ointments and creams. Ointments are semisolidpreparations which are typically based on petrolatum or other petroleumderivatives. Creams containing the selected active agent, are typicallyviscous liquid or semisolid emulsions, often either oil-in-water orwater-in-oil. Cream bases are typically water-washable, and contain anoil phase, an emulsifier and an aqueous phase. The oil phase, alsosometimes called the “internal” phase, is generally comprised ofpetrolatum and a fatty alcohol such as cetyl or stearyl alcohol; theaqueous phase usually, although not necessarily, exceeds the oil phasein volume, and generally contains a humectant. The emulsifier in a creamformulation is generally a nonionic, anionic, cationic or amphotericsurfactant. The specific ointment or cream base to be used, as will beappreciated by those skilled in the art, is one that will provide foroptimum drug delivery. As with other carriers or vehicles, an ointmentbase should be inert, stable, nonirritating and nonsensitizing.

Unlike typical peptide formulations, the peptides of this inventioncomprising D-form amino acids can be administered, even orally, withoutprotection against proteolysis by stomach acid, etc. Nevertheless, incertain embodiments, peptide delivery can be enhanced by the use ofprotective excipients. This is typically accomplished either bycomplexing the polypeptide with a composition to render it resistant toacidic and enzymatic hydrolysis or by packaging the polypeptide in anappropriately resistant carrier such as a liposome. Means of protectingpolypeptides for oral delivery are well known in the art (see, e.g.,U.S. Pat. No. 5,391,377 describing lipid compositions for oral deliveryof therapeutic agents).

Elevated serum half-life can be maintained by the use ofsustained-release protein “packaging” systems. Such sustained releasesystems are well known to those of skill in the art. In one preferredembodiment, the ProLease biodegradable microsphere delivery system forproteins and peptides (Tracy (1998) Biotechnol. Prog., 14: 108; Johnsonet al. (1996) Nature Med. 2: 795; Herbert et al. (1998), Pharmaceut.Res. 15, 357) a dry powder composed of biodegradable polymericmicrospheres containing the active agent in a polymer matrix that can becompounded as a dry formulation with or without other agents.

The ProLease microsphere fabrication process was specifically designedto achieve a high encapsulation efficiency while maintaining integrityof the active agent. The process consists of (i) preparation offreeze-dried drug particles from bulk by spray freeze-drying the drugsolution with stabilizing excipients, (ii) preparation of a drug-polymersuspension followed by sonication or homogenization to reduce the drugparticle size, (iii) production of frozen drug-polymer microspheres byatomization into liquid nitrogen, (iv) extraction of the polymer solventwith ethanol, and (v) filtration and vacuum drying to produce the finaldry-powder product. The resulting powder contains the solid form of theactive agents, which is homogeneously and rigidly dispersed withinporous polymer particles. The polymer most commonly used in the process,poly(lactide-co-glycolide) (PLG), is both biocompatible andbiodegradable.

Encapsulation can be achieved at low temperatures (e.g., −40° C.).During encapsulation, the protein is maintained in the solid state inthe absence of water, thus minimizing water-induced conformationalmobility of the protein, preventing protein degradation reactions thatinclude water as a reactant, and avoiding organic-aqueous interfaceswhere proteins may undergo denaturation. A preferred process usessolvents in which most proteins are insoluble, thus yielding highencapsulation efficiencies (e.g., greater than 95%).

In another embodiment, one or more components of the solution can beprovided as a “concentrate”, e.g., in a storage container (e.g., in apremeasured volume) ready for dilution, or in a soluble capsule readyfor addition to a volume of water.

The foregoing formulations and administration methods are intended to beillustrative and not limiting. It will be appreciated that, using theteaching provided herein, other suitable formulations and modes ofadministration can be readily devised.

B) Lipid-Based Formulations.

In certain embodiments, the active agents of this invention areadministered in conjunction with one or more lipids. The lipids can beformulated as an excipient to protect and/or enhance transport/uptake ofthe active agents or they can be administered separately.

Without being bound by a particular theory, it was discovered of thisinvention that administration (e.g. oral administration) of certainphospholipids can significantly increase HDL/LDL ratios. In addition, itis believed that certain medium-length phospholipids are transported bya process different than that involved in general lipid transport. Thus,co-administration of certain medium-length phospholipids with the activeagents of this invention confer a number of advantages: They protect theactive agents from digestion or hydrolysis, they improve uptake, andthey improve HDL/LDL ratios.

The lipids can be formed into liposomes that encapsulate the activeagents of this invention and/or they can be complexed/admixed with theactive agents and/or they can be covalently coupled to the activeagents. Methods of making liposomes and encapsulating reagents are wellknown to those of skill in the art (see, e.g., Martin andPapahadjopoulos (1982) J. Biol. Chem., 257: 286-288; Papahadjopoulos etal. (1991) Proc. Natl. Acad. Sci. USA, 88: 11460-11464; Huang et al.(1992) Cancer Res., 52:6774-6781; Lasic et al. (1992) FEBS Lett., 312:255-258, and the like).

Preferred phospholipids for use in these methods have fatty acidsranging from about 4 carbons to about 24 carbons in the sn-1 and sn-2positions. In certain preferred embodiments, the fatty acids aresaturated. In other preferred embodiments, the fatty acids can beunsaturated. Various preferred fatty acids are illustrated in Table 19.TABLE 19 Suitable fatty acids in the sn-1 and/or sn-2 position of thepreferred phospholipids for administration of active agents describedherein. Carbon No. Common Name IUPAC Name  3:0 Propionoyl Trianoic  4:0Butanoyl Tetranoic  5:0 Pentanoyl Pentanoic  6:0 Caproyl Hexanoic  7:0Heptanoyl Heptanoic  8:0 Capryloyl Octanoic  9:0 Nonanoyl Nonanoic 10:0Capryl Decanoic 11:0 Undcanoyl Undecanoic 12:0 Lauroyl Dodecanoic 13:0Tridecanoyl Tridecanoic 14:0 Myristoyl Tetradecanoic 15:0 PentadecanoylPentadecanoic 16:0 Palmitoyl Hexadecanoic 17:0 HeptadecanoylHeptadecanoic 18:0 Stearoyl Octadecanoic 19:0 Nonadecanoyl Nonadecanoic20:0 Arachidoyl Eicosanoic 21:0 Heniecosanoyl Heniecosanoic 22:0Behenoyl Docosanoic 23:0 Trucisanoyl Trocosanoic 24:0 LignoceroylTetracosanoic 14:1 Myristoleoyl (9-cis) 14:1 Myristelaidoyl (9-trans)16:1 Palmitoleoyl (9-cis) 16:1 Palmitelaidoyl (9-trans)The fatty acids in these positions can be the same or different.Particularly preferred phospholipids have phosphorylcholine at the sn-3position.

C) Specialized Delivery/Devices.

1. Drug-Eluting Stents.

Restenosis, the reclosure of a previously stenosed and subsequentlydilated peripheral or coronary vessel occurs at a significant rate(e.g., 20-50% for these procedures) and is dependent on a number ofclinical and morphological variables. Restenosis may begin shortlyfollowing an angioplasty procedure, but usually ceases at the end ofapproximately six (6) months.

A recent technology that has been developed to address the problem ofrestenosis in intravascular stents. Stents are typically devices thatare permanently implanted (expanded) in coronary and peripheral vessels.The goal of these stents is to provide a long-term “scaffolding” orsupport for the diseased (stenosed) vessels. The theory being, if thevessel is supported from the inside, it will not close down orrestenose.

Known stent designs include, but are not limited to monofilament wirecoil stents (see, e.g., U.S. Pat. No. 4,969,458 which is incorporatedherein by reference); welded metal cages (see, e.g., U.S. Pat. Nos.4,733,665 and 4,776,337 which are incorporated herein by reference),thin-walled metal cylinders with axial slots formed around thecircumference (see, e.g., U.S. Pat. Nos. 4,733,665, 4,739,762, 4,776,337which are incorporated herein by reference). Known constructionmaterials for use in stents include, but are not limited to polymers,organic fabrics and biocompatible metals, such as, stainless steel,gold, silver, tantalum, titanium, and shape memory alloys such asNitinol.

To further prevent restenosis, stents can be covered and/or impregnatedwith one or more pharmaceutical, e.g., in controlled releaseformulations to inhibit cell proliferation associated with rest enosis.Most commonly such “drug-eluting” stents are designed to deliver variouscancer drugs (cytotoxins).

However, because of their activity in mitigating inflammatory responses,reducing and/or eliminated oxidized lipids and/or other oxidizedspecies, inhibiting macrophage chemotactic activity and the like, theactive agents described herein are well suited to prevent restenosis.Thus, in certain embodiments, this invention contemplates stents havingone or more of the active agents described herein coated on the surfaceand/or retained within cavities or microcavities in the surface of thestent.

In certain embodiments the active agents are contained withinbiocompatible matrices (e.g. biocompatible polymers such as urethane,silicone, and the like). Suitable biocompatible materials are described,for example, in U.S. Patent Publications 2005/0084515, 2005/00791991,2005/0070996, and the like which are incorporated herein by reference.In various embodiments the polymers include, but are not limited tosilicone-urethane copolymer, a polyurethane, a phenoxy, ethylene vinylacetate, polycaprolactone, poly(lactide-co-glycolide), polylactide,polysulfone, elastin, fibrin, collagen, chondroitin sulfate, abiocompatible polymer, a biostable polymer, a biodegradable polymer

Thus, in certain embodiments this invention provides a stent fordelivering drugs to a vessel in a body. The stent typically comprisesstent framework including a plurality of reservoirs formed therein. Thereservoirs typically include an active agent and/or activeagent-containing polymer positioned in the reservoir and/or coated onthe surface of the stent. In various embodiments the stent is a metallicbase or a polymeric base. Certain preferred stent materials include, butare not limited to stainless steel, nitinol, tantalum, MP35N alloy,platinum, titanium, a suitable biocompatible alloy, a suitablebiocompatible polymer, and/or a combination thereof.

In various embodiments where the stent comprises pores (e.g.reservoirs), the pores can include micropores (e.g., having a diameterthat ranges from about 10 to about 50 μm, preferably about 20 μm orless). In various embodiments the micropores have a depth in the rangeof about 10 μm to about 50 μm. In various embodiments the microporesextend through the stent framework having an opening on an interiorsurface of the stent and an opening on an exterior surface of the stent.In certain embodiments the stent can, optionally comprise a cap layerdisposed on the interior surface of the stent framework, the cap layercovering at least a portion of the through-holes and providing a barriercharacteristic to control an elution rate of the active agent(s) in thepolymer from the interior surface of the stent framework. In variousembodiments the reservoirs comprise channels along an exterior surfaceof the stent framework. The stent can optionally have multiple layers ofpolymer where different layers of polymer carry different activeagent(s) and/or other drugs.

In certain embodiments the stent comprises: an adhesion layer positionedbetween the stent framework and the polymer. Suitable adhesion layersinclude, but are not limited to a polyurethane, a phenoxy,poly(lactide-co-glycolide)-, polylactide, polysulfone, polycaprolactone,an adhesion promoter, and/or a combination thereof.

In addition to stents, the active agents can be coated on or containedwithin essentially any implantable medical device configured forimplantation in a extravascular and/or intravascular location.

Also provided are methods of manufacturing a drug-polymer stent,comprising. The methods involve providing a stent framework; cutting aplurality of reservoirs in the stent framework, e.g., using a high powerlaser; applying one or more of the active agents and/or a drug polymerto at least one reservoir; drying the drug polymer; applying a polymerlayer to the dried drug polymer; and drying the polymer layer. Theactive agent(s) and/or polymer(s) can be applied by any convenientmethod including but not limited to spraying, dipping, painting,brushing and dispensing.

Also provided are methods of treating a vascular condition and/or acondition characterized by an inflammatory response and/or a conditioncharacterized by the formation of oxidized reactive species. The methodstypically involve positioning a stent or other implantable device asdescribed above within the body (e.g. within a vessel of a body) andeluting at least active agent from at least one surface of the implant.

2. Impregnated Grafts and Transplants.

Vascular grafts can be classified as either biological or synthetic.There are two commonly used types of biological grafts. An autograft isone taken from another site in the patient. In peripheral vascularsurgery by far the most commonly used such graft is the long saphenousvein. This can be used in situ with the valves surgically destroyed withan intraluminal cutting valvutome.

Alternatively, the vein can be removed and reversed but this typicallyproduces a discrepancy between the anastomotic size of the artery andvein. In thoracic surgery the use of internal mammary artery forcoronary artery bypass surgery is another example of an autograft. Anallograft is one taken from another animal of the same species.Externally supported umbilical vein is rarely used but is an example ofsuch a graft.

Synthetic grafts are most commonly made from Dacron orpolytetrafluoroethylene (PTFE). Dacron grafts are frequently used inaortic and aorto-iliac surgery. Below the inguinal ligament the resultsof all synthetic grafts are inferior to those obtained with the use ofvein grafts. Suitable vein is not always available and in this situationPTFE is typically used. It can be used in conjunction with vein as acomposite graft. Neointimal hyperplasia at the distal anastomosis can bereduced by the incorporation of a segment of vein as either a MillarCuff or Taylor Patch to improve the long-term patency of the grafts.

The commonest complications associated with the use of vascular graftsinclude Graft occlusion, Graft infection, true and false aneurysms atthe site of anastomosis, distal embolization, and erosion in to adjacentstructures—e.g. Aorto-enteric fistulae. Many of these conditions areassociated with an inflammatory response, macrophage migration into thesite, and/or the formation of reactive oxygen species (e.g., oxidizedlipids). To reduce such complications, the graft (synthetic orbiological can be soaked, or otherwise coated, with one or more of theactive agents described herein.

In addition, it is contemplated that other implantable tissues ormaterials can similarly be impregnated or coated with one or more activeagents of this invention. Thus, for example, in certain embodiments thisinvention contemplates the use of impregnated sutures to minimizeinflammation and/or infection and/or tissue rejection.

3. Subcutaneous Matrices.

In certain embodiments, one or more active agents described herein areadministered alone or in combination with other therapeutics asdescribed herein in implantable (e.g., subcutaneous) matrices.

A major problem with standard drug dosing is that typical delivery ofdrugs results in a quick burst of medication at the time of dosing,followed by a rapid loss of the drug from the body. Most of the sideeffects of a drug occur during the burst phase of its release into thebloodstream. Secondly, the time the drug is in the bloodstream attherapeutic levels is very short, most is used and cleared during theshort burst.

Drugs (e.g., the active agents described herein) imbedded in variousmatrix materials for sustained release provides some solution to theseproblems. Drugs embedded, for example, in polymer beads or in polymerwafers have several advantages. First, most systems allow slow releaseof the drug, thus creating a continuous dosing of the body with smalllevels of drug. This typically prevents side effects associated withhigh burst levels of normal injected or pill based drugs. Secondly,since these polymers can be made to release over hours to months, thetherapeutic span of the drug is markedly increased. Often, by mixingdifferent ratios of the same polymer components, polymers of differentdegradation rates can be made, allowing remarkable flexibility dependingon the agent being used. A long rate of drug release is beneficial forpeople who might have trouble staying on regular dosage, such as theelderly, but is also an ease of use improvement that everyone canappreciate. Most polymers can be made to degrade and be cleared by thebody over time, so they will not remain in the body after thetherapeutic interval.

Another advantage of polymer based drug delivery is that the polymersoften can stabilize or solubilize proteins, peptides, and other largemolecules that would otherwise be unusable as medications. Finally, manydrug/polymer mixes can be placed directly in the disease area, allowingspecific targeting of the medication where it is needed without losingdrug to the “first pass” effect. This is certainly effective fortreating the brain, which is often deprived of medicines that can'tpenetrate the blood/brain barrier.

A number of implantable matrix (sustained release) systems are know tothose of skill and can readily be adapted for use with one or more ofthe active agents described herein. Suitable sustained release systemsinclude, but are not limited to Re-Gel®, SQ2Gel®, and Oligosphere® byMacroMed, ProLease® and Medisorb® by Alkermes, Paclimer® and Gliadel®Wafer by Guilford pharmaceuticals, the Duros implant by Alza, acousticbioSpheres by Point Biomedical, the Intelsite capsule by Scintipharma,Inc., and the like.

4. Other “Specialty Delivery Systems”.

Other “specialty” delivery systems include, but are not limited to lipidbased oral mist that allows absorption of drugs across the oral mucosa,developed by Generex Biotechnology, the oral transmucosal system (OTS™)by Anesta Corp., the inhalable dry powder and PulmoSpheres technology byInhale Therapeutics, the AERx® Pulmonary Drug Delivery System byAradigm, the AIR mechanism by Alkermes, and the like.

Another approach to delivery developed by Alkermes is a system targetedfor elderly and pediatric use, two populations for which taking pills isoften difficult is known as Drug Sipping Technology (DST). Themedication is placed in a drinking straw device, prevented from fallingout by filters on either end of it. The patient merely has to drinkclear liquid (water, juice, soda) through the straw. The drug dissolvesin the liquid as it is pulled through and is ingested by the patient.The filter rises to the top of the straw when all of the medication istaken. This method has the advantage in that it is easy to use, theliquid often masks the medication's taste, and the drug is pre-dissolvedfor more efficient absorption.

It is noted that these uses and delivery systems are intended to beillustrative and not limiting. Using the teachings provided herein,other uses and delivery systems will be known to those of skill in theart.

VI. Additional Pharmacologically Active Agents.

Combined Active Agents

In various embodiments, the use of combinations of two or more activeagents described is contemplated in the treatment of the variouspathologies/indications described herein. The use of combinations ofactive agents can alter pharmacological activity, bioavailability, andthe like.

By way of illustration, it is noted that D-4F rapidly associates withpre-beta HDL and HDL and then is rapidly cleared from the circulation(it is essentially non-detectable 6 hours after an oral dose), whileD-[113-122]apoJ slowly associates with pre-beta HDL and to a lesserextent with HDL but remains associated with these HDL fractions for atleast 36 hours. FREL associates with HDL and only HDL but remainsdetectable in HDL for much longer than D-4F (i.e., it is detectable inHDL 48 hours after a single oral dose in mice). In certain embodimentsthis invention thus contemplates combinations of, for example, thesethree peptides to reduce the amount to reduce production expense, and/orto optimize dosage regimen, therapeutic profile, and the like. Incertain embodiments combinations of the active agents described hereincan be simply coadministered and/or added together to form a singlepharmaceutical formulation. In certain embodiments the various activeagent(s) can be complexed together (e.g. via hydrogen bonding) to formactive agent complexes that are more effective than the parent agents.

Use with Additional Pharmacologically Active Materials.

Additional pharmacologically active materials (i.e., drugs) can bedelivered in conjunction with one or more of the active agents describedherein. In certain embodiments, such agents include, but are not limitedto agents that reduce the risk of atherosclerotic events and/orcomplications thereof. Such agents include, but are not limited to betablockers, beta blockers and thiazide diuretic combinations, statins,aspirin, ace inhibitors, ace receptor inhibitors (ARBs), and the like.

It was discovered that, adding a low dosage active agent (e.g., of D-4F)(1 μg/ml) to the drinking water of apoE null mice for 24 hours did notsignificantly improve HDL function (see, e.g., related application U.S.Ser. No. 10/423,830, filed on Apr. 25, 2003, which is incorporatedherein by reference). In addition, adding 0.05 mg/ml of atorvastatin orpravastatin alone to the drinking water of the apoE null mice for 24hours did not improve HDL function. However, when D-4F1 μg/ml was addedto the drinking water together with 0.05 mg/ml of atorvastatin orpravastatin there was a significant improvement in HDL function). Indeedthe pro-inflammatory apoE null HDL became as anti-inflammatory as 350μg/ml of normal human HDL (h, HDL see, e.g., related application U.S.Ser. No. 10/423,830).

Thus, doses of D-4F alone, or statins alone, which by themselves had noeffect on HDL function when given together acted synergistically. WhenD-4F and a statin were given together to apo E null mice, theirpro-inflammatory HDL at 50 μg/ml of HDL-cholesterol became as effectiveas normal human HDL at 350 μg/ml of HDL-cholesterol in preventing theinflammatory response induced by the action of HPODE oxidizing PAPC incocultures of human artery wall cells.

Thus, in certain embodiments this invention provides methods forenhancing the activity of statins. The methods generally involveadministering one or more of the active agents described herein, asdescribed herein in conjunction with one or more statins. The activeagents achieve synergistic action between the statin and the agent(s) toameliorate one or more symptoms of atherosclerosis. In this contextstatins can be administered at significantly lower dosages therebyavoiding various harmful side effects (e.g., muscle wasting) associatedwith high dosage statin use and/or the anti-inflammatory properties ofstatins at any given dose are significantly enhanced.

Suitable statins include, but are not limited to pravastatin(Pravachol/Bristol-Myers Squibb), simvastatin (Zocor/Merck), lovastatin(Mevacor/Merck), and the like.

In various embodiments the active agent(s) described herein areadministered in conjunction with one or more beta blockers. Suitablebeta blockers include, but are not limited to cardioselective (selectivebeta 1 blockers), e.g., acebutolol (Sectral™), atenolol (Tenormin™),betaxolol (Kerlone™), bisoprolol (Zebeta™), metoprolol (Lopressor™), andthe like. Suitable non-selective blockers (block beta 1 and beta 2equally) include, but are not limited to carteolol (Cartrol™), nadolol(Corgard™), penbutolol (Levatol™), pindolol (Visken™), propranolol(Inderal™), timolol (Blockadren™), labetalol (Normodyne™, Trandate™),and the like.

Suitable beta blocker thiazide diuretic combinations include, but arenot limited to Lopressor HCT, ZIAC, Tenoretic, Corzide, Timolide,Inderal LA 40/25, Inderide, Normozide, and the like.

Suitable ace inhibitors include, but are not limited to captopril (e.g.Capoten™ by Squibb), benazepril (e.g., Lotensin™ by Novartis), enalapril(e.g., Vasotec™ by Merck), fosinopril (e.g., Monopril™ byBristol-Myers), lisinopril (e.g. Prinivil™ by Merck or Zestril™ byAstra-Zeneca), quinapril (e.g. Accupril™ by Parke-Davis), ramipril(e.g., Altace™ by Hoechst Marion Roussel, King Pharmaceuticals),imidapril, perindopril erbumine (e.g., Aceon™ by Rhone-Polenc Rorer),trandolapril (e.g., Mavik™ by Knoll Pharmaceutical), and the like.Suitable ARBS (Ace Receptor Blockers) include but are not limited tolosartan (e.g. Cozaar™ by Merck), irbesartan (e.g., Avapro™ by Sanofi),candesartan (e.g., Atacand™ by Astra Merck), valsartan (e.g., Diovan™ byNovartis), and the like.

In various embodiments, one or more agents described herein areadministered with one or more of the drugs identified below.

Thus, in certain embodiments one or more active agents are administeredin conjunction with cholesteryl ester transfer protein (CETP) inhibitors(e.g., torcetrapib, JTT-705. CP-529414) and/or acyl-CoA:cholesterolO-acyltransferase (ACAT) inhibitors (e.g., Avasimibe (CI-1011), CP113818, F-1394, and the like), and/or immunomodulators (e.g., FTY720(sphingosine-1-phosphate receptor agonist), Thalomid (thalidomide),Imuran (azathioprine), Copaxone (glatiramer acetate), Certican®(everolimus), Neoral® (cyclosporine), and the like), and/ordipeptidyl-peptidase-4 (DPP4) inhibitors (e.g.,2-Pyrrolidinecarbonitrile,1-[[[2-[(5-cyano-2-pyridinyl)amino]ethyl]amino]acetyl], see also U.S.Patent Publication 2005-0070530), and/or calcium channel blockers (e.g.,Adalat, Adalat CC, Calan, Calan SR, Cardene, Cardizem, Cardizem CD,Cardizem SR, Dilacor-XR, DynaCirc, Isoptin, Isoptin SR, Nimotop,Norvasc, Plendil, Procardia, Procardia XL, Vascor, Verelan), and/orperoxisome proliferator-activated receptor (PPAR) agonists for, e.g., α,γ; δ receptors (e.g., Azelaoyl PAF, 2-Bromohexadecanoic acid,Ciglitizone, Clofibrate, 15-Deoxy-δ^(12,14)-prostaglandin J₂,Fenofibrate, Fmoc-Leu-OH, GW1929, GW7647,8(S)-Hydroxy-(5Z,9E,11Z,14Z)-eicosatetraenoic acid (8(S)-HETE),Leukotriene B₄, LY-171,883 (Tomelukast), Prostaglandin A₂, ProstaglandinJ₂, Tetradecylthioacetic acid (TTA), Troglitazone (CS-045), WY-14643(Pirinixic acid)), and the like.

In certain embodiments one or more of the active agents are administeredin conjunction with fibrates (e.g., clofibrate (atromid), gemfibrozil(lopid), fenofibrate (tricor), etc.), bile acid sequestrants (e.g.,cholestyramine, colestipol, etc.), cholesterol absorption blockers(e.g., ezetimibe (Zetia), etc.), Vytorin ((ezetimibe/simvastatincombination), and/or steroids, warfarin, and/or aspirin, and/or Bcr-Ablinhibitors/antagonists (e.g., Gleevec (Imatinib Mesylate), AMN107,STI571 (CGP57148B), ON 012380, PLX225, and the like), and/or reninangiotensin pathway blockers (e.g., Losartan (Cozaar®), Valsartan(Diovan®), Irbesartan (Avapro®), Candesartan (Atacand®), and the like),and/or angiotensin II receptor antagonists (e.g., losartan (Cozaar),valsartan (Diovan), irbesartan (Avapro), candesartan (Atacand) andtelmisartan (Micardis), etc.), and/or PKC inhibitors (e.g., CalphostinC, Chelerythrine chloride, Chelerythrine.chloride, Copperbis-3,5-diisopropylsalicylate, Ebselen, EGF Receptor (human) (651-658)(N-Myristoylated), Gö 6976, H-7. dihydrochloride,1-O-Hexadecyl-2-O-methyl-rac-glycerol, Hexadecyl-phosphocholine(C_(16:0)); Miltefosine, Hypericin, Melittin (natural), Melittin(synthetic), ML-7. hydrochloride, ML-9. hydrochloride,Palmitoyl-DL-camitine. hydrochloride, Protein Kinase C (19-31), ProteinKinase C (19-36), Quercetin.dihydrate, Quercetin.dihydrate,D-erythro-Sphingosine (isolated), D-erythro-Sphingosine (synthetic),Sphingosine, N,N-dimethyl, D-erythro-Sphingosine, Dihydro-,D-erythro-Sphingosine, N,N-Dimethyl-, D-erythro-Sphingosine chloride,N,N,N-Trimethyl-, Staurosporine, Bisindolylmaleimide I, G-6203, and thelike).

In certain embodiments, one or more of the active agents areadministered in conjunction with ApoAI, Apo A-I derivatives and/oragonists (e.g., ApoAI milano, see, e.g., U.S. Patent Publications20050004082, 20040224011, 20040198662, 20040181034, 20040122091,20040082548, 20040029807, 20030149094, 20030125559, 20030109442,20030065195, 20030008827, and 20020071862, and U.S. Pat. Nos. 6,831,105,6,790,953, 6,773,719, 6,713,507, 6,703,422, 6,699,910, 6,680,203,6,673,780, 6,646,170, 6,617,134, 6,559,284, 6,506,879, 6,506,799,6,459,003, 6,423,830, 6,410,802, 6,376,464, 6,367,479, 6,329,341,6,287,590, 6,090,921, 5,990,081, and the like), renin inhibitors (e.g.,SPP630 and SPP635, SPP100, Aliskiren, and the like), and/or MRantagonist (e.g., spironolactone, aldosterone glucuronide, and thelike), and/or aldosterone synthase inhibitors, and/or alpha-adrenergicantagonists (e.g., Aldomet® (Methyldopa), Cardura® (Doxazosin),Catapres®; Catapres-TTS®; Duraclon™ (Clonidine), Dibenzyline®(Phenoxybenzamine), Hylorel® (Guanadrel), Hytrin® (Terazosin),Minipress® (Prazosin), Tenex® (Guanfacine), Guanabenz, Phentolamine,Reserpine, and the like), and/or liver X receptor (LXR) agonists (e.g.,T0901317, GW3965, ATI-829, acetyl-podocarpic dimer (APD), and the like),and/or framesoid X receptor (FXR) agonists (e.g., GW4064,6alpha-ethyl-chenodeoxycholic acid (6-ECDCA), T0901317, and the like),and/or plasminogen activator-1 (PAI-1) inhibitors (see, e.g.,oxime-based PAI-1 inhibitors, see also U.S. Pat. No. 5,639,726, and thelike), and/or low molecular weight heparin, and/or AGEinhibitors/breakers (e.g., Benfotiamine, aminoguanidine, pyridoxamine,Tenilsetam, Pimagedine, and the like) and/or ADP receptor blockers(e.g., Clopidigrel, AZD6140, and the like), and/or ABCA1 agonists,and/or scavenger receptor B1 agonists, and/or Adiponectic receptoragonist or adiponectin inducers, and/or stearoyl-CoA Desaturase I (SCD1)inhibitors, and/or Cholesterol synthesis inhibitors (non-statins),and/or Diacylglycerol Acyltransferase I (DGAT1) inhibitors, and/orAcetyl CoA Carboxylase 2 inhibitors, and/or LP-PLA2 inhibitors, and/orGLP-1, and/or glucokinase activator, and/or CB-1 agonists, and/oranti-thrombotic/coagulants, and/or Factor Xa inhibitors, and/orGPIIb/IIIa inhibitors, and/or Factor VIIa inhibitors, and/or Tissuefactor inhibitors, and/or anti-inflammatory drugs, and/or Probucol andderivatives (e.g. AGI-1067, etc.), and/or CCR2 antagonists, and/orCX3CR1 antagonists, and/or IL-1 antagonists, and/or nitrates and NOdonors, and/or phosphodiesterase inhibitors, and the like.

IX. Kits for the Treatment of One or More Indications.

In another embodiment this invention provides kits for amelioration ofone or more symptoms of atherosclerosis or for the prophylactictreatment of a subject (human or animal) at risk for atherosclerosisand/or the treatment or prophylaxis of one or more of the conditionsdescribed herein. The kits preferably comprise a container containingone or more of the active agents described herein. The active agent(s)can be provided in a unit dosage formulation (e.g. suppository, tablet,caplet, patch, etc.) and/or may be optionally combined with one or morepharmaceutically acceptable excipients.

The kit can, optionally, further comprise one or more other agents usedin the treatment of the condition/pathology of interest. Such agentsinclude, but are not limited to, beta blockers, vasodilators, aspirin,statins, ace inhibitors or ace receptor inhibitors (ARBs) and the like,e.g. as described above.

In addition, the kits optionally include labeling and/or instructionalmaterials providing directions (i.e., protocols) for the practice of themethods or use of the “therapeutics” or “prophylactics” of thisinvention. Preferred instructional materials describe the use of one ormore active agent(s) of this invention to mitigate one or more symptomsof atherosclerosis (or other pathologies described herein) and/or toprevent the onset or increase of one or more of such symptoms in anindividual at risk for atherosclerosis (or other pathologies describedherein). The instructional materials may also, optionally, teachpreferred dosages/therapeutic regiment, counter indications and thelike.

While the instructional materials typically comprise written or printedmaterials they are not limited to such. Any medium capable of storingsuch instructions and communicating them to an end user is contemplatedby this invention. Such media include, but are not limited to electronicstorage media (e.g., magnetic discs, tapes, cartridges, chips), opticalmedia (e.g., CD ROM), and the like. Such media may include addresses tointernet sites that provide such instructional materials.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Example 1 Use of ApoJ-Related Peptides to Mediate Symptoms ofAtherosclerosis

Prevention of LDL-Induced Monocyte Chemotactic Activity

FIG. 1 illustrates a comparison of the effect of D-4F (Anantharamaiah etal. (2002) Circulation, 105: 290-292) with the effect of an apoJ peptidemade from D amino acids (D-J336,Ac-L-L-E-Q-L-N-E-Q-F-N-W-V-S-R-L-A-N-L-T-Q-G-E-NH₂, SEQ ID NO: 1177)) onthe prevention of LDL-induced monocyte chemotactic activity in vitro ina co-incubation. Human aortic endothelial cells were incubated withmedium alone (no addition), with control human LDL (200 μg protein/ml)or control human LDL+control human HDL (350 μg HDL protein/ml). D-J336or D-4F was added to other wells in a concentration range as indicatedplus control human LDL (200 μg protein/ml). Following overnightincubation, the supernatants were assayed for monocyte chemotacticactivity. As shown in FIG. 1, the in vitro concentration of the apoJvariant peptide that prevents LDL-induced monocyte chemotactic activityby human artery wall cells is 10 to 25 times less than the concentrationrequired for the D-4F peptide.

Prevention of LDL-Induced Monocyte Chemotactic Activity by Pre-Treatmentof Artery Wall Cells with D-J336

FIG. 2 illustrates a comparison of the effect of D-4F with the effect ofD-J336 on the prevention of LDL induced monocyte chemotactic activity ina pre-incubation. Human aortic endothelial cells were pre-incubated withD-J336 or D-4F at 4, 2, and 1 μg/ml for DJ336 or 100, 50, 25, and 12.5μg/ml for D-4F for 6 hrs. The cultures were then washed and wereincubated with medium alone (no addition), or with control human LDL(200 μg protein/ml), or with control human LDL+control human HDL (350 μgHDL protein/ml) as assay controls. The wells that were pre-treated withpeptides received the control human LDL at 200 μg protein/ml. Followingovernight incubation, the supernatants were assayed for monocytechemotactic activity.

As illustrated in FIG. 2, the ApoJ variant peptide was 10-25 times morepotent in preventing LDL oxidation by artery wall cells in vitro.

The Effect of apo J Peptide Mimetics on HDL Protective Capacity in LDLReceptor Null Mice.

D-4F designated as F, or the apoJ peptide made from D amino acids(D-J336, designated as J) was added to the drinking water of LDLreceptor null mice (4 per group) at 0.25 or 0.5 mg per ml of drinkingwater. After 24- or 48-hrs blood was collected from the mice and theirHDL was isolated and tested for its ability to protect againstLDL-induced monocyte chemotactic activity. Assay controls includedculture wells that received no lipoproteins (no addition), or controlhuman LDL alone (designated as LDL, 200 μg cholesterol/ml), or controlLDL+control human HDL (designated as +HDL, 350 μg HDL cholesterol). Fortesting the mouse HDL, the control LDL was added together with mouse HDL(+F HDL or +J HDL) to artery wall cell cultures. The mouse HDL was addedat 100 μg cholesterol/ml respectively. After treatment with either D-4For D-J336 the mouse HDL at 100 μg/ml was as active as 350 μg/ml ofcontrol human HDL in preventing the control LDL from inducing the arterywall cells to produce monocyte chemotactic activity. The reason for thediscrepancy between the relative doses required for the D-J336 peptiderelative to D-4F in vitro and in vivo may be related to the solubilityof the peptides in water and we believe that when measures are taken toachieve equal solubility the D-J peptides will be much more active invivo as they are in vitro.

Protection Against LDL-Induced Monocyte Chemotactic Activity by HDL fromapo E Null Mice Given Oral Peptides.

FIG. 4 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on HDL protective capacity. ApoE null mice (4 per group) wereprovided with D-4F (designated as F) at 50, 30, 20, 10, 5 μg per ml ofdrinking water or apoJ peptide (designated as J) at 50, 30 or 20 μg perml of drinking water. After 24 hrs blood was collected, plasmafractionated by FPLC and fractions containing LDL (designated as mLDLfor murine LDL) and fractions containing HDL (designated as mHDL) wereseparately pooled and HDL protective capacity against LDL oxidation asdetermined by LDL-induced monocyte chemotactic activity was determined.For the assay controls the culture wells received no lipoproteins (noadditions), mLDL alone (at 200 μg cholesterol/ml), or mLDL+standardnormal human HDL (designated as Cont. h HDL, at 350 μg HDLcholesterol/ml).

For testing the murine HDL, mLDL together with murine HDL (+F mHDL or +JmHDL) were added to artery wall cell cultures. The HDL from the micethat did not receive any peptide in their drinking water is designatedas no peptide mHDL. The murine HDL was used at 100 μg cholesterol/ml.After receiving D-4F or D-J336 the murine HDL at 100 μg/ml was as activeas 350 μg/ml of normal human HDL. As shown in FIG. 4, when added to thedrinking water the D-J peptide was as potent as D-4F in enhancing HDLprotective capacity in apo E null mice.

Ability of LDL Obtained from apoE Null Mice Given Oral Peptides toInduce Monocyte Chemotactic Activity.

FIG. 5 illustrates the effect of oral apo A-I peptide mimetic and apoJpeptide on LDL susceptibility to oxidation. ApoE null mice (4 per group)were provided, in their drinking water, with D-4F (designated as F) at50, 30, 20, 10, 5 μg per ml of drinking water or the apoJ peptide(D-J336 made from D amino acids and designated as J) at 50, 30 or 20 μgper ml of drinking water. After 24 hrs blood was collected from the miceshown in FIG. 4, plasma fractionated by FPLC and fractions containingLDL (designated as mLDL for murine LDL) were pooled and LDLsusceptibility to oxidation as determined by induction of monocytechemotactic activity was determined. For the assay controls the culturewells received no lipoproteins (no additions), mLDL alone (at 200 μgcholesterol/ml), or mLDL+standard normal human HDL (designated as Cont.h HDL, 350 μg HDL cholesterol).

Murine LDL, mLDL, from mice that received the D-4F (F mLDL) or thosethat received the apoJ peptide (J mLDL) were added to artery wall cellcultures. LDL from mice that did not receive any peptide in theirdrinking water is designated as No peptide LDL.

As shown in FIG. 5, when added to the drinking water, D-J336 wasslightly more potent than D-4F in rendering the LDL from apo E null miceresistant to oxidation by human artery wall cells as determined by theinduction of monocyte chemotactic activity.

Protection Against Phospholipid Oxidation and Induction of MonocyteChemotactic Activity by HDL obtained from apo E Null Mice Given OralPeptides.

FIG. 6 illustrates the effect of oral apoA-1 peptide mimetic and apoJpeptide on HDL protective capacity. ApoE null mice (4 per group) wereprovided with D-4F (designated as F) at 50, 30, 20, 10, 5 μg per ml ofdrinking water or apoJ peptide (D-J336 made from D amino acids anddesignated as J) at 50, 30 or 20 μg per ml of drinking water. After 24hrs blood was collected, plasma fractionated by FPLC and fractionscontaining HDL (designated as mHDL) were pooled and HDL protectivecapacity against PAPC oxidation as determined by the induction ofmonocyte chemotactic activity was determined. For the assay controls theculture wells received no lipoproteins (no additions), the phospholipidPAPC at 20 μg/ml+HPODE, at 1.0 μg/ml, or PAPC+HPODE plus standard normalhuman HDL (at 350 μg HDL cholesterol/ml and designated as +Cont. h HDL).

For testing the murine HDL, PAPC+HPODE together with murine HDL (+F mHDLor +J mHDL) were added to artery wall cell cultures. The HDL from micethat did not receive any peptide in their drinking water is designatedas “no peptide mHDL”. The murine HDL was used at 100 μg cholesterol/ml.

The data show in FIG. 6 indicate that, when added to the drinking water,D-J336 was as potent as D-4F in causing HDL to inhibit the oxidation ofa phospholipid PAPC by the oxidant HPODE in a human artery wallco-culture as measured by the generation of monocyte chemotacticactivity

Effect of Oral apoA-1 Peptide Mimetic and apoJ Peptide on PlasmaParaoxonase Activity in Mice.

FIG. 7 shows the effect of oral apoA-1 peptide mimetic and apoJ peptideon plasma paraoxonase activity in mice. ApoE null mice (4 per group)were provided with D-4F designated as F at 50, 10, 5 or 0 μg per ml ofdrinking water or apoJ peptide (D-J336 made from D amino acids anddesignated as J) at 50, 10 or 5 μg per ml of drinking water. After 24hrs blood was collected and plasma was assayed for PON1 activity. Thesedata demonstrate that, when added to the drinking water, D-J336 was atleast as potent as D-4F in increasing the paraoxonase activity of apo Enull mice.

Example 2 Oral G* Peptides Increase HDL Protective Capacity in Apo EDeficient Mice

Female, 4 month old apoE deficient mice (n=4 per group) were treatedwith G* peptides having the following amino acid sequences. Peptide113-122=Ac-L V G R Q L E E F L-NH₂ (SEQ ID NO. 9), Peptide 336-357=Ac-LL E Q L N E Q F N W V S R L A N L T Q G E-NH₂ (SEQ ID NO. 17), andPeptide 377-390=Ac-P S G V T E V V V K L F D S-NH₂ (SEQ ID NO. 19).

Each mouse received 200 μg of the peptide by stomach tube. Four hourslater blood was obtained, plasma separated, lipoproteins fractionatedand HDL (at 25 μg per ml) was assayed for protective capacity againstthe oxidation of LDL (at 100 μg per ml) in cultures of human artery wallcells. The data are shown in FIG. 8. The peptide afforded significantHDL-protective capacity in the mice.

In another experiment, female, 4 month old apoE deficient mice (n=4 pergroup) were treated with the 11 amino acid G* peptide 146-156 with thesequence: Ac-Q Q T H M L D V M Q D-NH₂. (SEQ ID NO:11). The micereceived the peptide in their drinking water at the indicatedconcentrations (see FIG. 9). Following eighteen hrs, blood was obtained,plasma separated, lipoproteins fractionated and HDL (at 50 μgcholesterol per ml) was assayed for protective capacity against theoxidation of PAPC (at 25 μg per ml)+HPODE (at 1.0 μg per ml) in culturesof human artery wall cells. Assay controls included No additions,PAPC+HPODE and PAPC+HPODE plus Control HDL (designated as +HDL). Thedata are mean +/−SD of the number of migrated monocytes in nine highpower fields in triplicate cultures. Asterisks indicate significance atthe level of p<0.05 vs. the water control (0 μg/ml).

Example 3 Solution Phase Chemistry for Peptide Synthesis

In certain embodiments, a solution-phase synthesis chemistry provides amore economical means of synthesizing peptides of this invention.

Prior to this invention synthesis was typically performed using anall-solid phase synthesis chemistry. The solid phase synthesis ofpeptides of less than 9 amino acids is much more economical than thesolid phase synthesis of peptides of more than 9 amino acids. Synthesisof peptides of more than 9 amino acids results in a significant loss ofmaterial due to the physical dissociation of the elongating amino acidchain from the resin. The solid phase synthesis of peptides containingless than 9 amino acids is much more economical because the there isrelatively little loss of the elongating chain from the resin.

In certain embodiments, the solution phase synthesis functions byconverting the synthesis of the 18 amino acid apoA-I mimetic peptide, 4F(and other related peptides) from an all solid phase synthesis to eitheran all solution phase synthesis or to a combination of solid phasesynthesis of three chains each containing, e.g., 6 amino acids followedby the assembly of the three chains in solution. This provides a muchmore economical overall synthesis. This procedure is readily modifiedwhere the peptides are not 18 amino acids in length. Thus, for example,a 15 mer can be synthesized by solid phase synthesis of three 5 mersfollowed by assembly of the three chains in solution. A 14 mer can besynthesized by the solid phase synthesis of two 5 mers and one 4 merfollowed by assembly of these chains in solution, and so forth.

A) Summary of Synthesis Protocol.

An scheme for the synthesis of the peptide D4F(Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH₂, (SEQ ID NO:5) isillustrated in Table 20. (The scheme and yields for the synthesis areshown in Table 20. TABLE 20 Illustrative solution phase synthesisscheme. Final Wt. of Pure Wt. of Wt. of Crude Peptide Fmoc CouplingResin Peptide (gms) (mg) Synthesis Resin Amino Acid Reagent (gms) Yield(%) Yield ((%) Methods Used for D4F Synthesis Stepwise Rink Amide 6Equiv HBTU/ 4 2.0 500 Solid Phase (1 mmole) HOBT 1.8 gms 86 25 StepwiseRink Amide 2 Equiv DIC/HOBT 3.9 2.0 450 Solid Phase (1 mmole) 1.8 gms 8622.5 Fragment Rink Amide HBTU/ 3.3 1.0 100 coupling (1 mmole) HOBT (6 +6 + 6) 1.8 gms* 43 10 Synthesis of D4F Fragments Fragment 1 (2HN-KFKEAF(SEQ ID NO: 1178) on rink amide resin (K and E are properly protected)Fragment 2 Cl-TrT-Resin 6 Equiv HBTU/ 11 2.2 6 residues (5 mmol) HOBTcrude stepwise 6.5 gms protected Solid Phase 36Fmoc-Y(But)-D(But)-K(Boc)-V-A-E(But)-COOH (SEQ ID NO: 1179) Fragment 2Cl-TrT-Resin 6 Equiv HBTU/ 10 1.8 6 residues (5 mmol) HOBT crudestepwise 6.5 gms protected Solid Phase 32 Ac-D(But)-W-F-K(Boc)-A-F-COOH(SEQ ID NO: 1180)Synthesis by solution phase using fragments produced by the solid phasemethod.Fragment Wang resin. C-terminal hexapeptide (subjected to ammonolysis).Yield quantitative.1. NH2-K(Boc)-F-K(Boc)-E(But)-A-F-Wang resin (SEQ ID NO: 1181)NH2-K(Boc)-F-K(Boc)-E(But)-A-F-CO-NH2 (SEQ ID NO: 1182)Fragment 2 from above was coupled to fragment 1 in DMF using DIC/HOBT.Fmoc-Y(But)-D(But)-K(Bpc)-V-A-E(But)-K(Boc)-F-K(Boc)-E(But)-F-Co-NH2(SEQ ID NO: 1183) 12 residue peptide was characterized as free peptideafter removing protecting groups. Yield was 50%Fmoc from the above-12 rtesidue was removed by piperidine in DMF (20%.After drying the peptide was copled to Fragment 3 using DCl/HOBT in DMF.Ac-D(But)-W-F-K(Boc)-A-F-Y(But)-D(but)-K(Boc)-V-A-E(But)-K(Boc)-F-K(Boc)-E(But)-A-FCO-NH2(SEQ ID NO: 1184)Protected peptide yield was quantitative.Protecting groups removed using mixture of TFA (80%), phenol (5%),thioanisole (5%). water) 5%), triisopropylsilane (TIS, 5%), stirred for90 min.Precipitated by ether and purified by C-4 HPLC column. Yield 25%

B) Details of Synthesis Protocol.

1. Fragment Condensation Procedure to Synthesize D-4F

Fragments synthesized for fragment condensation on solid phase are:

-   -   Fragment 1: Ac-D(OBut)-W-F-K(εBoc)-A-F-COOH (SEQ ID NO:1185);    -   Fragment 2:Fmoc-Y(OBut)-D(OBut)-K(εBoc)-V-A-E(OBut)-COOH (SEQ ID        NO:1186); and    -   Fragment 3 Fmoc-K(εBoc)F-K(εBoc)-E(OBut)-A-F- Rink amide resin        (SEQ ID NO:1187).

Fragment 1 was left on the resin to obtain final peptide amide after TFAtreatment.

To synthesize fragment 1: Fmoc-Phe (1.2 equivalents) was added tochlorotrityl resin (Nova Biochem, 1.3 mMol/g substitution, 5 mMol or 6.5g was used) in presence of six equivalents of DIEA inDMF:dichloromethane (1:1)) and stirred for 4 h. Excess of functionalityon the resin was capped with methanol in presence of dichloromethane andDIEA. After the removal of Fmoc-Fmoc amino acid derivatives (2equivalents) were added using HOBt/HBTU reagents as described above.Final Fmoc-D(OBut)-W-F-K(εBoc)-A-F Chlorotrityl resin was treated withFmoc deblocking agent and acetylated with 6 equivalents of aceticanhydride in presence of diisoprolylethyl amine. The resultingAc-D(OBut)-W-F-K(εBoc)-A-F-resin was treated with a mixture oftrifluoroethanol-acetic acid-dichloromethane (2:2:6, 10 ml/g of resin)for 4 h at room temperature. After removal of the resin by filtration,the solvent was removed by aziotropic distillation with n-hexane undervacuum. The residue (1.8 g) was determined by mass spectral analysis tobe Ac-D(OBut)-W-F-K(εBoc)-A-F-COOH (SEQ ID NO:1188).

Fragment 2, Fmoc-Y(OBut)-D(OBut)-K(εBoc)-V-A-E(OBut)-COOH (SEQ IDNO:1189), was obtained using the procedure described for Fragment 1.Final yield was 2.2 g.

Fragment 3. 0.9 g (0.5 mmol) of Rink amide resin (Nova Biochem) was usedto obtain fragment Rink amide resin was treated with 20% piperidine indichloromethane for 5 min once and 15 min the second time (Fmocdeblocking reagents). 1. 2 equivalents of Fmoc-Phe was condensed usingcondensing agents HOBt/HBTU (2 equivalents in presence of few drops ofdiisopropylethyl amine) (amino acid condensation). Deblocking andcondensation of the rest of the amino acids were continued to obtain theof Fmoc-K(εBoc)F-K(εBoc)-E(OBut)-A-F-rink amide resin (SEQ ID NO:1190).Fmoc was cleaved and the peptide resin K(εBoc)F-K(εBoc)-E(OBut)-A-F-rink amide resin (SEQ ID NO:1190) was used for fragment condensation asdescribed below.

Fragment 2 in DMF was added to Fragment 3 (1.2 equivalents) usingHOBt-HBTU procedure in presence of DIEA overnight. After washing theresin with DMF and deblocking Fmoc-Fragment 1 (1.2 equivalents) wasadded to the dodecapeptide resin using HOBt-HBTU procedure overnight.

The final peptide resin (3.3 g) was treated with a mixture ofTFA-Phenol-triisopropylsilane-thioanisole-water (80:5:5:5) for 1.5 h (10ml of the reagent/g of the resin). The resin was filtered off and thesolution was diluted with 10 volumes of ether. Precipitated peptide wasisolated by centrifugation and washed twice with ether. 1 g of the crudepeptide was subjected to HPLC purification to obtain 100 mg of thepeptide.

2. Characterization of Peptide.

The peptide was identified by mass spectral and analytical HPLC methods.

As shown in FIG. 14 the product of the solution phase synthesis schemeis very biologically active in producing HDL and pre-beta HDL thatinhibit LDL-induced monocyte chemotaxis in apo E null mice. ApoE nullmice were fed 5 micrograms of the D-4F synthesized as described above(Frgmnt) or the mice were given the same amount of mouse chow withoutD-4F (Chow). Twelve hours after the feeding was started, the mice werebled and their plasma was fractionated on FPLC. LDL (100 microgramsLDL-cholesterol) was added to cocultures of human artery wall cellsalone (LDL) or with a control human HDL (Control HDL) or with HDL (50micrograms HDL-cholesterol) or post-HDL (pHDL; prebeta HDL) from micethat did (Frgmnt) or did not (Chow) receive the D-4F and the monocytechemotactic activity produced was determined

Example 4 Comparison of D-4F and Reverse (Retro-) D-4F Activity

As shown in FIG. 16, the biological activities of D-4F and reverse RD-4Fare not significantly different. Female apoE null mice were administeredby stomach tube 0, 3, 6, 12, or 25 micrograms of D-4F or Reverse D-4F in100 microliters of water. Blood was obtained 7 hours later and theplasma was fractionated by FPLC. A standard control human LDL was addedto human artery wall cells at a concentration of 100 micrograms ofLDL-cholesterol/mL (LDL). The resulting monocyte chemotactic activitywas normalized to 1.0. The same LDL at the same concentration was addedto the human artery wall cells together with HDL at 50 microgramsHDL-cholesterol/mL from a normal human (hHDL) or from the apoE null micethat received the dose of D-4F or Reverse D-4F shown on the X-axis. Theresulting monocyte chemotactic activity was normalized to that of theLDL added without HDL. The resulting value is the HDL InflammatoryIndex. The results shown are the Mean ±S.D. for the data from threeseparate experiments.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

1: A peptide that ameliorates a symptom of atherosclerosis, wherein saidpeptide comprises the amino acid sequence or the retro amino acidsequence of a peptide listed in Table 2, Table 3, Table, Table 5, Table6, Table 7, Table 8, Table 9, or Table
 10. 2: The peptide of claim 1,wherein said peptide further comprises a protecting group coupled to theamino or carboxyl terminus. 3: The peptide of claim 1, wherein saidpeptide further comprises a first protecting group coupled to the aminoterminus and a second protecting group coupled to the carboxyl terminus.4: The peptide of claim 2, wherein said protecting groups areindependently selected from the group consisting of acetyl, amide, and 3to 20 carbon alkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group,1-fluorenecarboxylic group, 9-florenecarboxylic group,9-fluorenone-1-carboxylic group, benzyloxycarbonyl, Xanthyl (Xan),Trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt),4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr), Mesitylene-2-sulphonyl(Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl (Tos), 2,2,5,7,8-pentamethylchroman-6-sulphonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl(MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl), Benzoyl (Bz),3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z),2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).5: The peptide of claim 1, wherein one or more amino acids comprisingsaid peptide are “D” amino acids. 6: The peptide of claim 1, wherein allamino acids comprising said peptide “D” amino acids. 7: The peptide ofclaim 1, wherein said peptide is mixed with a pharmacologicallyacceptable excipient. 8: The peptide of claim 1, wherein said peptide ismixed with a pharmacologically acceptable excipient suitable for oraladministration to a mammal. 9: The peptide of claims 2-8, wherein saidpeptide comprises a protecting group coupled to the amino terminal andsaid amino terminal protecting group is a protecting group selected fromthe group consisting of acetyl, propeonyl, and a 3 to 20 carbon alkyl.10: The peptide of claim 9, wherein said peptide comprises a protectinggroup coupled to the carboxyl terminal and said carboxyl terminalprotecting group is an amide. 11: The peptide of claim 9, wherein saidpeptide comprises: a first protecting group coupled to the aminoterminus wherein said protecting group is a protecting group selectedfrom the group consisting of acetyl, propeonyl, and a 3 to 20 carbonalkyl; and a second protecting group coupled to the carboxyl terminaland said carboxyl terminal protecting group is an amide. 12: A peptidethat ameliorates a symptom of atherosclerosis, wherein said peptide:consists of 18 amino acids, said 18 amino acids consisting of 3 alanines(A), 2 aspartates (D), 2 glutamates (E), 4 phenylalanines (F), 4 lysines(K), 1 valine (V), 1 tryptophan (W), and 1 tyrosine (Y); wherein saidpeptide forms a class A amphipathic helix; comprises at least one “D”amino acid residue; and protects a phospholipid against oxidation by anoxidizing agent. 13: The peptide of claim 12, wherein said peptide hasthe amino acid sequence or the retro amino acid sequence of a peptidelisted in Table
 4. 14: The peptide of claim 12, wherein said peptidefurther comprises a protecting group coupled to the amino or carboxylterminus. 15: The peptide of claim 12, wherein said peptide furthercomprises a first protecting group coupled to the amino terminus and asecond protecting group coupled to the carboxyl terminus. 16: Thepeptide of claim 14, wherein said protecting groups are independentlyselected from the group consisting of acetyl, amide, and 3 to 20 carbonalkyl groups, Fmoc, Tboc, 9-fluoreneacetyl group, 1-fluorenecarboxylicgroup, 9-florenecarboxylic group, 9-fluorenone-1-carboxylic group,benzyloxycarbonyl, Xanthyl (Xan), Trityl (Trt), 4-methyltrityl (Mtt),4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulphonyl (Mtr),Mesitylene-2-sulphonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), Tosyl(Tos), 2,2,5,7,8-pentamethyl chroman-6-sulphonyl (Pmc), 4-methylbenzyl(MeBzl), 4-methoxybenzyl (MeOBzl), Benzyloxy (BzlO), Benzyl (Bzl),Benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys),1-(4,4-dimentyl-2,6-diaxocyclohexylidene)ethyl (Dde), 2,6-dichlorobenzyl(2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z),2-bromobenzyloxycarbonyl (2-Br-Z), Benzyloxymethyl (Bom),t-butoxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butoxymethyl (Bum),t-butoxy (tBuO), t-Butyl (tBu), Acetyl (Ac), and Trifluoroacetyl (TFA).17: The peptide of claim 12, wherein all enantiomeric amino acids are“D” amino acids. 18-24. (canceled) 25: A peptide that ameliorates asymptom of atherosclerosis, wherein said peptide: ranges in length fromabout 18 to 37 amino acids and comprises at least 3 alanines (A), 2aspartates (D), 2 glutamates (E), 4 phenylalanines (F), 4 lysines (K), 1valine (V), 1 tryptophan (W), 1 tyrosine (Y); wherein said peptide formsa class A amphipathic helix; comprises at least one “D” amino acidresidue; and protects a phospholipid against oxidation by an oxidizingagent. 26: The peptide of claim 25, wherein said peptide furthercomprises a protecting group coupled to the amino or carboxyl terminus.27-31. (canceled) 32: The peptide of claim 25, wherein said peptidecomprises an amino acid sequence selected from the group consisting ofD-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F (SEQ ID NO: 1191),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1192),-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F-P-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-F-F(SEQ ID NO: 1193),-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F—P-D-W-F-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F(SEQ ID NO: 1194),D-K-L-K-A-F-Y-D-K-V-F-E-W-A-K-E-A-F-P-D-K-L-K-A-F-Y-D-K-V-F-E-W-L-K-E-A-F(SEQ ID NO: 1195),D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L-P-D-K-W-K-A-V-Y-D-K-F-A-E-A-F-K-E-F-L(SEQ ID NO: 1196),D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-P-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-(SEQ ID NO: 1197), or the reverse of any of these sequences. 33-37.(canceled) 38: A peptide that forms a class A amphipathic helix or aclass Y amphipathic helix and is described by the formula:D¹-X¹-X²-K¹-Y¹-X³-X⁴-D²-K²-X⁵-Y-D³-K³-X⁶-K⁴-D⁴-Y²-X⁷ claim wherein: X¹,X², X³, X⁴, X⁵, and X⁶ are independently selected from the groupconsisting of Leu, norLeu, Val, Ile, Trp, Phe, Tyr, β-Nal, and α-Nal,and all X residues are on the non-polar face of said peptide, except forone that can be on the polar face between two K residues; K¹, K², K³,and K⁴ are independently Lys or Arg, and no more than two K's areadjacent to each other in a helical wheel diagram of said peptide; Y¹and Y² are independently selected from the group consisting of Ala, His,Ser, Gln, Asn, and Thr, when present on the non-polar face of saidmolecule; when one of Y¹ or Y² are present on the polar face of saidmolecule, the Y¹ or Y² on the polar face of said molecule is selectedfrom the group consisting of Ala, His, Ser, Gln, Asn, and Thr; D¹, D²,D³, and D⁴ are independently Asp or Glu, and no more than 3 Ds arecontiguous in a helical wheel diagram of said peptide, and the remainingD is separated from the other D's by a Y. 39: The peptide of claim 64,wherein said peptide comprises the amino acid sequence or the retroamino acid sequence of a peptide listed in Table
 5. 40-49. (canceled)50: A method of treating a vascular condition and/or a conditioncharacterized by an inflammatory response and/or a conditioncharacterized by the formation of oxidized reactive species in a mammal,said method comprising: administering to a mammal in need thereof one ormore of the active agents described in Tables 2-18, and/or a smallorganic molecule as described herein in an amount sufficient toameliorate one or more symptoms of said condition. 51: The method ofclaim 50, wherein said active agent is a polypeptide comprising theamino acid sequence of 4F (SEQ ID NO:5).
 52. (canceled) 53: The methodof claim 50, wherein said active agent is administered in conjunctionwith a drug selected from the group consisting of CETP inhibitors,FTY720, Certican, DPP4 inhibitors, Calcium channel blockers, ApoA1derivative or mimetic or agonist, PPAR agonists, Steroids, Gleevec,Cholesterol Absorption blockers (Zetia), Vytorin, Any Renin Angiotensinpathway blockers, Angiotensin II receptor antagonist (Diovan etc), ACEinhibitors, Renin inhibitors, MR antagonist and Aldosterone synthaseinhibitor, Beta-blockers, Alpha-adrenergic antagonists, LXR agonist, FXRagonist, Scavenger Receptor B1 agonist, ABCA1 agonist, Adiponecticreceptor agonist or adiponectin inducers, Stearoyl-CoA Desaturase I(SCD1) inhibitor, Cholesterol synthesis inhibitors (non-statins),Diacylglycerol Acyltransferase I (DGAT1) inhibitor, Acetyl CoACarboxylase 2 inhibitor, PAI-1 inhibitor, LP-PLA2 inhibitor, GLP-1,Glucokinase activator, CB-1 agonist, AGE inhibitoribreaker, PKCinhibitors, Anti-thrombotic/coagulants: Aspirin, ADP receptor blockerse.g. Clopidigrel, Factor Xa inhibitor, GPIIb/IIIa inhibitor, Factor VIIainhibitor, Warfarin, Low molecular weight heparin, Tissue factorinhibitor, Anti-inflammatory drugs: Probucol and derivative e.g.AGI-1067 etc, CCR2 antagonist, CX3CR1 antagonist, IL-1 antagonist,Nitrates and NO donors, and Phosphodiesterase inhibitors. 54: A methodof ameliorating one or more symptoms of a condition selected from thegroup consisting of atherosclerotic plaque formation, atheroscleroticlesion formation, myocardial infarction, stroke, congestive heartfailure, arteriole function, arteriolar disease, arteriolar diseaseassociated with aging, arteriolar disease associated with Alzheimer'sdisease, arteriolar disease associated with chronic kidney disease,arteriolar disease associated with hypertension, arteriolar diseaseassociated with multi-infarct dementia, arteriolar disease associatedwith subarachnoid hemorrhage, peripheral vascular disease, chronicobstructive pulmonary disease (COPD), emphysema, asthma, idiopathicpulmonary fibrosis, pulmonary fibrosis, adult respiratory distresssyndrome, osteoporosis, Paget's disease, coronary calcification,rheumatoid arthritis, polyarteritis nodosa, polymyalgia rheumatica,lupus erythematosus, multiple sclerosis, Wegener's granulomatosis,central nervous system vasculitis (CNSV), Sjögren's syndrome,scleroderma, polymyositis, AIDS inflammatory response, bacterialinfection, fungal infection, viral infection, parasitic infection,influenza, avian flu, viral pneumonia, endotoxic shock syndrome, sepsis,sepsis syndrome, trauma/wound, organ transplant, transplantatherosclerosis, transplant rejection, corneal ulcer,chronic/non-healing wound, ulcerative colitis, reperfusion injury(prevent and/or treat), ischemic reperfusion injury (prevent and/ortreat), spinal cord injuries (mitigating effects), cancers,myeloma/multiple myeloma, ovarian cancer, breast cancer, colon cancer,bone cancer osteoarthritis, inflammatory bowel disease, allergicrhinitis, cachexia, diabetes, Alzheimer's disease, implanted prosthesis,biofilm formation, Crohns' disease, dermatitis, acute and chronic,eczema, psoriasis, contact dermatitis, scleroderma, Type I Diabetes,Type II Diabetes, juvenile onset diabetes, prevention of the onset ofdiabetes, diabetic nephropathy, diabetic neuropathy, diabeticretinopathy, erectile dysfunction, macular degeneration, multiplesclerosis, nephropathy, neuropathy, Parkinson's Disease, peripheralvascular disease, and meningitis, said method comprising administeringto a mammal in need thereof an active agent described in Tables 2-18,and/or a small organic molecule as described herein in an amountsufficient ameliorate a symptom of said condition.
 55. (canceled) 56: Astent for delivering drugs to a vessel in a body comprising: a stentframework including a plurality of reservoirs formed therein, and apeptide comprising the amino acid sequence or the retro amino acidsequence of a peptide listed in Tables 2-18. 57-73. (canceled) 74: Amethod of manufacturing a drug-polymer stent, comprising: providing astent framework; cutting a plurality of reservoirs in the stentframework; applying a composition comprising one or more peptidescomprising the amino acid sequence or the retro amino acid sequence of apeptide listed in Table 4, Table 5, or Table 6 to at least onereservoir; and drying the composition.
 75. (canceled) 76: A method oftreating a vascular condition, comprising: positioning a stent accordingto claim 56 within a vessel of a body; expanding the stent; and elutingat least one active agent from at least a surface of the stent.